Miracle-dz/UniFAR-FTUs huggingface dataset:
Characterization and Molecular Cloning of a Novel Enzyme, Inorganic Polyphosphate/ATP-Glucomannokinase, of Arthrobacter sp. Strain KM

ABSTRACT A bacterium exhibiting activities of several inorganic polyphosphate [poly(P)]- and ATP-dependent kinases, including glucokinase, NAD kinase, mannokinase, and fructokinase, was isolated, determined to belong to the genus Arthrobacter, and designated Arthrobacter sp. strain KM. Among the kinases, a novel enzyme responsible for the poly(P)- and ATP-dependent mannokinase activities was purified 2,200-fold to homogeneity from a cell extract of the bacterium. The purified enzyme was a monomer with a molecular mass of 30 kDa. This enzyme phosphorylated glucose and mannose with a high affinity for glucose, utilizing poly(P) as well as ATP, and was designated poly(P)/ATP-glucomannokinase. The Km values of the enzyme for glucose, mannose, ATP, and hexametaphosphate were determined to be 0.50, 15, 0.20, and 0.02 mM, respectively. The catalytic sites for poly(P)-dependent phosphorylation and ATP-dependent phosphorylation of the enzyme were found to be shared, and the poly(P)-utilizing mechanism of the enzyme was shown to be nonprocessive. The gene encoding the poly(P)/ATP-glucomannokinase was cloned from Arthrobacter sp. strain KM, and its nucleotide sequence was determined. This gene contained an open reading frame consisting of 804 bp coding for a putative polypeptide with a calculated molecular mass of 29,480 Da. The deduced amino acid sequence of the polypeptide exhibited homology to the amino acid sequences of the poly(P)/ATP-glucokinase of Mycobacterium tuberculosis H37Rv (level of homology, 45%), ATP-dependent glucokinases of Corynebacterium glutamicum (45%), Renibacterium salmoninarum (45%), and Bacillus subtilis (35%), and proteins of bacteria belonging to the order Actinomyces whose functions are not known. Alignment of these homologous proteins revealed seven conserved regions. The mannose and poly(P) binding sites of poly(P)/ATP-glucomannokinase are discussed.
Spinal Anesthesia with Bupivacaine Decreases Cerebral Blood Flow in Former Preterm Infants

Spinal anesthesia is commonly used in former preterm infants (FPI). In these patients, hypotension induced by spinal anesthesia may impair cerebral blood flow. We measured cerebral blood flow velocity (CBFV) by transcranial Doppler ultrasound to assess the effect of hypotension induced by spinal anesthesia on cerebral hemodynamics. Twelve FPI scheduled for inguinal hernia repair were operated under spinal anesthesia using 1 mg/kg isobaric 0.5% bupivacaine. Systolic, diastolic, and mean middle cerebral artery CBFV were measured at 5 min before and 5 min and 10 min after spinal anesthesia using a transcranial pulsed Doppler ultrasonography. Arterial blood pressure and heart rate were recorded simultaneously. Cerebral arteries resistance index (RI) was calculated as RI = (peak systolic CBFV − end-diastolic CBFV)/peak systolic CBFV. Diastolic CBFV decreased significantly from 30.0 ± 11.1 cm/s to 20.1 ± 8.4 cm/s at 5 min and to 20.1 ± 7.0 cm/s at 10 min. RI increased significantly from 0.7 ± 0.1 to 0.8 ± 0.1 at 5 min and 10 min. Systolic, diastolic, and mean arterial blood pressures decreased significantly at the same time intervals. We suggest that in FPI, spinal anesthesia induces a decrease in cerebral blood flow related to changes in arterial blood pressure. Whether these changes have deleterious consequences remains to be determined.
Analysis of single EBER‐positive and negative tumour cells in EBV‐harbouring B‐cell non‐Hodgkin lymphomas

In situ hybridization for Epstein–Barr virus (EBV) encoded small nuclear RNAs (EBERs) is the method of choice for the detection of EBV infection at the single cell level. With the application of this technique it was shown that non‐Hodgkin's lymphomas of B‐cell type may be associated with an EBV infection of tumour cells. Interestingly, in many EBV‐positive cases, only a proportion of the tumour cell population has been found to be EBER‐positive. To clarify whether EBV is absent or whether EBER gene expression is downregulated in EBER‐negative tumour cells, single EBER‐positive and negative tumour cells were isolated from paraffin sections from four B‐cell non‐Hodgkin's lymphoma cases with partial EBER expression in the neoplastic cells. These single cells were then screened for the presence of EBV DNA by a nested single cell PCR. EBV DNA was undetectable in all but one of the 86 EBER‐negative cells, whereas in the EBER‐positive cells EBV‐specific DNA amplificates could be generated following single cell PCR. This finding prompts the conclusion that the inability to detect EBERs in a proportion of tumour cells is not due to a down‐regulation of gene expression but to a real absence of EBV from these cells. This partial absence of EBV is thought to be caused by a loss of the EBV episomes during cell division, rather than by infection of only a proportion of the tumour cells. Copyright © 2001 John Wiley & Sons, Ltd.
The effect of green tea polyphenols on macrophage migration inhibitory factor‐associated steroid resistance

Background Recently, evidence has been obtained to suggest that inflammation is provoked through upregulation of macrophage migration inhibitory factor (MIF) expression by steroids. However, little is known regarding the effect of steroids on MIF expression in human keratinocytes and the counter‐effect of epigallocatechin‐3‐gallate (EGCG), a member of the class of green tea polyphenols.
Sequential regimen of chemotherapy, reduced-intensity conditioning for allogeneic stem-cell transplantation, and prophylactic donor lymphocyte transfusion in high-risk acute myeloid leukemia and myelodysplastic syndrome.

PURPOSE To improve the effect of allogeneic stem-cell transplantation by sequential use of intensive chemotherapy, reduced-intensity conditioning (RIC), and prophylactic donor lymphocyte transfusions (pDLTs) in high-risk acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). PATIENTS AND METHODS In a prospective study of 75 consecutive patients (median age, 52.3 years), high risk was defined by progressive or refractory disease (n = 59), second remission after early relapse (n = 8), or first remission with poor prognosis based on cytogenetics or delayed response to induction therapy (n = 8). Unfavorable karyotypes were found in 49% of informative patients, and 68 patients had medical contraindications against standard conditioning. Fludarabine (30 mg/m2), cytarabine (2 g/m2), and amsacrine (100 mg/m2) for 4 days were used for cytoreduction. After 3 days of rest, RIC consisted of 4 Gy total-body irradiation, antithymocyte globulin, and 80 to 120 mg/kg cyclophosphamide. Thirty-one patients had an HLA-identical sibling donor; 44 patients had an unrelated and/or HLA-mismatched donor. pDLT was given from day +120 in patients who were not receiving immunosuppression and were free of graft-versus-host disease (GvHD). RESULTS Complete remission was induced in 66 patients (88%). With a median follow-up of 35.1 months (range, 13.6 to 47.6 months), 2-year overall and leukemia-free survival were 42% and 40%, respectively. Outcome of patients with refractory disease or with complex cytogenetic aberrations was identical to that of better prognostic subgroups. Survival was best in patients who received high CD34+ cell numbers, and in patients with limited GvHD. CONCLUSION Sequential use of intensive chemotherapy, RIC transplantation, and pDLT represents a promising approach to the treatment of high-risk AML and MDS, particularly in patients with most unfavorable prognoses.
A protein that interacts with members of the nuclear hormone receptor family: identification and cDNA cloning.

In search of proteins which interact with activated steroid hormone receptors, we screened a human liver lambda gt11 expression library with the glucocorticoid receptor. We identified and cloned a cDNA sequence of 1322 bp that encodes a protein of 274 aa. This protein consists predominantly of hydrophilic amino acids and contains a putative bipartite nuclear localization signal. The in vitro translated receptor-associating protein runs in SDS/polyacrylamide gels with an apparent molecular mass of 46 kDa. By use of the bacterially expressed fusion protein with glutathione S-transferase we have found that interaction is not limited to the glucocorticoid receptor but included other nuclear receptors--most notably, the estrogen and thyroid receptors. Binding also occurs with the glucocorticoid receptor complexed with the antiglucocorticoid RU 38486, with the estrogen receptor complexed with the antiestrogen 4-hydroxytamoxifen or ICI 164,384, and even with receptors not complexed with ligand. Association with steroid hormone receptors depends on prior receptor activation--i.e., release from heat shock proteins. The sequence identified here appears to be a general partner protein for nuclear hormone receptors, with the gene being expressed in a variety of mammalian tissues.
Plasmodium falciparum Heterochromatin Protein 1 Marks Genomic Loci Linked to Phenotypic Variation of Exported Virulence Factors

Epigenetic processes are the main conductors of phenotypic variation in eukaryotes. The malaria parasite Plasmodium falciparum employs antigenic variation of the major surface antigen PfEMP1, encoded by 60 var genes, to evade acquired immune responses. Antigenic variation of PfEMP1 occurs through in situ switches in mono-allelic var gene transcription, which is PfSIR2-dependent and associated with the presence of repressive H3K9me3 marks at silenced loci. Here, we show that P. falciparum heterochromatin protein 1 (PfHP1) binds specifically to H3K9me3 but not to other repressive histone methyl marks. Based on nuclear fractionation and detailed immuno-localization assays, PfHP1 constitutes a major component of heterochromatin in perinuclear chromosome end clusters. High-resolution genome-wide chromatin immuno-precipitation demonstrates the striking association of PfHP1 with virulence gene arrays in subtelomeric and chromosome-internal islands and a high correlation with previously mapped H3K9me3 marks. These include not only var genes, but also the majority of P. falciparum lineage-specific gene families coding for exported proteins involved in host–parasite interactions. In addition, we identified a number of PfHP1-bound genes that were not enriched in H3K9me3, many of which code for proteins expressed during invasion or at different life cycle stages. Interestingly, PfHP1 is absent from centromeric regions, implying important differences in centromere biology between P. falciparum and its human host. Over-expression of PfHP1 results in an enhancement of variegated expression and highlights the presence of well-defined heterochromatic boundaries. In summary, we identify PfHP1 as a major effector of virulence gene silencing and phenotypic variation. Our results are instrumental for our understanding of this widely used survival strategy in unicellular pathogens.
Centennial Decline in the Trophic Level of an Endangered Seabird after Fisheries Decline

Abstract: Coastal marine ecosystems worldwide have undergone such profound transformations from overfishing that trophic interactions observed today might be artifacts of these changes. We determined whether the trophic level of an endangered seabird, the Marbled Murrelet (Brachyramphus marmoratus), has declined over the past 100 years after the collapse of Pacific sardine (Sardinops sadax) fisheries in the late 1940s and the recent declines of similar fisheries in central California. We compared stable‐isotope signatures of δ15N and δ13C in feathers of museum specimens collected before fisheries decline with values in murrelet feathers collected recently. Values of δ15N in prebreeding diets declined significantly, 1.4‰ or 38% of a trophic level, over the past century during cool ocean conditions and by 0.5‰ during warm conditions, whereas postbreeding values of δ15N were nearly constant. The δ13C values in prebreeding diets declined by 0.8‰, suggesting an increased importance of krill in modern compared with historic prebreeding diets, but postbreeding diets did not change. Stable‐isotope mixing models indicated that the proportion of energetically superior, high‐trophic‐level prey declined strongly whereas energetically poor, low‐trophic‐level and midtrophic‐level prey increased in the prebreeding diet in cool years when murrelet reproduction was likely to be high. Decreased prey resources have caused murrelets to fish further down on the food web, appear partly responsible for poor murrelet reproduction, and may have contributed to its listing under the U.S. Endangered Species Act.
Intraoperative arterial blood pressure lability is associated with improved 30 day survival.

BACKGROUND Arterial blood pressure lability, defined as rapid changes in arterial blood pressure, occurs commonly during anaesthesia. It is believed that hypertensive patients exhibit more lability during surgery and that lability is associated with poorer outcomes. Neither association has been rigorously tested. We hypothesized that hypertensive patients have more blood pressure lability and that increased lability is associated with increased 30 day mortality. METHODS This was a retrospective single-centre study of surgical patients from July 2008 to December 2012. Intraoperative data were extracted from the electronic anaesthesia record. Lability was calculated as the modulus of the percentage change in mean arterial pressure between consecutive 5 min intervals. The number of episodes of lability >10% was tabulated. Multivariate logistic regression was performed to determine the association between lability and 30 day mortality using derivation and validation cohorts. RESULTS Inclusion criteria were met by 52 919 subjects. Of the derivation cohort, 53% of subjects were hypertensive and 42% used an antihypertensive medication. The median number of episodes of lability >10% was 9 (interquartile range 5-14) per patient. Hypertensive subjects demonstrated more lability than normotensive patients, 10 (5-15) compared with 8 (5-12), P<0.0001. In subjects taking no antihypertensive medication, lability >10% was associated with decreased 30 day mortality, odds ratio (OR) per episode 0.95 [95% confidence interval (CI) 0.92-0.97], P<0.0001. This result was confirmed in the validation cohort, OR 0.96 (95% CI 0.93-0.99), P=0.01, and in hypertensive patients taking no antihypertensive medication, OR 0.96 (95% CI 0.93-0.99), P=0.002. Use of any antihypertensive medication class reduced this effect. CONCLUSIONS Intraoperative arterial blood pressure lability occurs more often in hypertensive patients. Contrary to common belief, increased lability was associated with decreased 30 day mortality.
Detection and care practices for postpartum depressive symptoms in public‐sector obstetric units in Mexico: Qualitative results from a resource‐constrained setting

BACKGROUND Postpartum depression (PPD) is amenable to detection and treatment, but effectively addressing it is contingent on policies, practices, and health care providers working together to address the issue. The aim of this study is to describe a sample of health care providers' existing practices in public-sector obstetric units in Mexico related to detecting and offering care to women with depressive symptomology. METHODS Semi-structured interviews with 40 health care providers (16 physicians, 13 nurses, three social workers, and eight psychologists) from one tertiary-level and two secondary-level, public-sector obstetric units, were conducted by members of a trained research team from May to July 2012. Qualitative data were analyzed in Spanish according to Grounded Theory, using Nvivo 10 software. RESULTS Lack of hospital guidelines, training, and time constraints are reasons given for not detecting PPD symptoms among women. Providers reported that their role is cursory and limited to giving women anticipatory guidance for what to expect emotionally after childbirth or providing a trusting atmosphere for women to express their feelings. Care is fragmented and inadequate, in part because of the lack of protocols that define who makes mental health referrals and where. Providers indicated PPD is important but not prioritized in health care for pregnant and postpartum women. CONCLUSION Critical needs in obstetric units include formal mental health care detection and care protocols during the perinatal period, strategies to address mental health needs despite short hospital stays, and training for providers on how to implement detection and care protocols and strategies.
Immediate loading of two unsplinted implants retaining the existing complete mandibular denture in elderly edentulous patients: 1-year results from a multicentre prospective cohort study.

PURPOSE To evaluate the outcome of two freestanding implants immediately loaded retaining an existing mandibular complete denture in elderly edentulous patients up to 1 year after loading. MATERIALS AND METHODS 42 patients with a mean age of 76.5 years (range 71 to 89) were selected for stabilising their mandibular complete denture with two implants placed mesial to the mandibular canine position bilaterally. Individual ball abutments were connected and torqued at 30 Ncm and the existing complete denture was immediately attached. No postoperative limitations to chewing function were given. The patients were evaluated clinically and radiographically at implant placement and at 6- and 12-month follow-up examinations. RESULTS At the 12-month follow-up no implant failed. Peri-implant bone resorption was 0.203 mm (CI 95% 0.322; 0.086) after 6 months and 0.298 mm (CI 95% 0.425; 0.173) after 12 months. Of the 42 cases, 3 had major prosthetic complications and 5 patients required minor extra maintenance appointments. CONCLUSIONS Within the limits of this study, it can be suggested that the immediate loading of two unsplinted implants retaining the existing complete mandibular denture in elderly patients can result in favourable implant survival and peri-implant bone healing, however larger and longer follow-ups of 5 years or more are needed.
Serum from Patients Undergoing Remote Ischemic Preconditioning Protects Cultured Human Intestinal Cells from Hypoxia-Induced Damage: Involvement of Matrixmetalloproteinase-2 and -9

Remote ischemic preconditioning (RIPC) can be induced by transient occlusion of blood flow to a limb with a blood pressure cuff and exerts multiorgan protection from ischemia/reperfusion injury. Ischemia/reperfusion injury in the intestinal tract leads to intestinal barrier dysfunction and can result in multiple organ failure. Here we used an intestinal cell line (CaCo-2) to evaluate the effects of RIPC-conditioned patient sera on hypoxia-induced cell damage in vitro and to identify serum factors that mediate RIPC effects. Patient sera (n = 10) derived before RIPC (T0), directly after RIPC (T1) and 1 h after RIPC (T2) were added to the culture medium at the onset of hypoxia until 48 h after hypoxia. Reverse transcription-polymerase chain reaction, lactate dehydrogenase (LDH) assays, caspase-3/7 assays, silver staining, gelatin zymography and Western blotting were performed. Hypoxia led to morphological signs of cell damage and increased the release of LDH in cultures containing sera T0 (P < 0.01) and T1 (P < 0.05), but not sera T2, which reduced the hypoxia-mediated LDH release compared with sera T0 (P < 0.05). Gelatin zymography revealed a significant reduction of activities of the matrixmetalloproteinase (MMP)-2 and MMP-9 in the protective sera T2 compared with the nonprotective sera T0 (MMP-2: P < 0.01; MMP-9: P < 0.05). Addition of human recombinant MMP-2 and MMP-9 to MMP-deficient culture media increased the sensitivity of CaCo-2 cells to hypoxia-induced cell damage (P < 0.05), but did not result in a reduced phosphorylation of prosurvival kinases p42/44 and protein kinase B (Akt) or increased activity of caspase-3/7. Our results suggest MMP-2 and MMP-9 as currently unknown humoral factors that may be involved in RIPC-mediated cytoprotection in the intestine.
Effects of drugs of abuse and cholinergic agents on delayed matching-to-sample responding in the squirrel monkey.

To study how drugs may disrupt short-term memory function, squirrel monkeys were trained to respond under a titrating delayed matching-to-sample schedule of reinforcement. Monkeys could respond on each of three keys in an operant chamber. At the start of each trial, the 30th response on the center key illuminated each of the side keys, one of which matched the stimulus presented on the center key (simultaneous matching). A response to the correct (matching) side key turned off all stimuli and initiated a delay, the length of which varied as a function of ongoing performance. After the delay, stimuli were randomly presented on two of the three keys. A response to the key which matched the color on the center key before the delay resulted in delivery of a food pellet (delayed matching). Incorrect simultaneous or delayed matching responses initiated a timeout. Under this procedure, diazepam and scopolamine decreased delayed matching accuracy at one or more doses that did not significantly decrease mean delay values, but only scopolamine decreased matching accuracy at a dose that did not significantly decrease response rates. Cocaine decreased mean delay values after the highest dose without affecting matching accuracy. Pentobarbital and methylscopolamine decreased matching accuracy and mean and maximum delay values after the highest doses. Nicotine and phencyclidine produced small decreases in delayed matching accuracy without affecting mean and maximum delay values. Caffeine, morphine, physostigmine and neostigmine did not alter matching performance even after doses that markedly decreased rates of responding. (ABSTRACT TRUNCATED AT 250 WORDS)
A Novel Archaeal Transcriptional Regulator of Heat Shock Response*

Archaea have a eukaryotic type of transcriptional machinery containing homologues of the transcription factors TATA-binding protein (TBP) and TFIIB (TFB) and a pol II type of RNA polymerase, whereas transcriptional regulators identified in archaeal genomes have bacterial counterparts. We describe here a novel regulator of heat shock response, Phr, from the hyperthermophilic archaeonPyrococcus furiosus that is conserved among Euryarchaeota. The protein specifically inhibited cell-free transcription of its own gene and from promoters of a small heat shock protein, Hsp20, and of an AAA+ ATPase. Inhibition of transcription was brought about by abrogating RNA polymerase recruitment to the TBP/TFB promoter complex. Phr bound to a 29-bp DNA sequence overlapping the transcription start site. Three sequences conserved in the binding sites of Phr, TTTA at −10, TGGTAA at the transcription start site, and AAAA at position +10, were required for Phr binding and are proposed as consensus regulatory sequences of Pyrococcus heat shock promoters. Shifting the growth temperature from 95 to 103 °C caused a dramatic increase of mRNA levels for theaaa+ atpase and phrgenes, but expression of the Phr protein was only weakly stimulated. Our findings suggest that heat shock response in Archaea is negatively regulated by a mechanism involving binding of Phr to conserved sequences.