Changes in myocardial substrate and energy metabolism by S-(4)-hydroxyphenylglycine and an N-(6)-derivative of adenosine

Summary In 15 mongrel open chest dogs oxidative myocardial carbohydrate utilization was stimulated by activation of pyruvatedehydrogenase with S-(4)-hydroxyphenylglycine (HPG) or by inhibition of lipolysis with N(6)-allyl-N(6)-cyclohexyladenosine (PAA). HPG and PAA shifted cardiac respiratory quotients (RQ) from 0.83 to 0.89 and 0.99, respectively. Oxygen extraction ratio of lactate was significantly increased by both interventions. Arterial nonesterified fatty acids (NEFA) concentration decreased significantly only by PAA. The oxygen saving potency of both interventions was quantified over a wide hemodynamic range by comparing the directly measured myocardial oxygen consumption (MVO₂) with the myocardial energy requirements calculated from its hemodynamic determinants according to the Bretschneider formula during base conditions and -stimulation. Inhibition of peripheral lipolysis with PAA reduced MVO₂ by 14 %, enzyme activation with HPG by 8 %. The results show that the efficiency of the myocardial energy supply can be influenced by manipulation of the oxidative substrate metabolism.Supported by the Deutsche Forschungsgemeinschaft Ka 577/1-1     

Functional motifs in the (6-4) photolyase crystal structure make a comparative framework for DNA repair photolyases and clock cryptochromes

Homologous flavoproteins from the photolyase (PHR)/cryptochrome (CRY) family use the FAD cofactor in PHRs to catalyze DNA repair and in CRYs to tune the circadian clock and control development. To help address how PHR/CRY members achieve these diverse functions, we determined the crystallographic structure of Arabidopsis thaliana (6-4) PHR (UVR3), which is strikingly (>65%) similar in sequence to human circadian clock CRYs. The structure reveals a substrate-binding cavity specific for the UV-induced DNA lesion, (6-4) photoproduct, and cofactor binding sites different from those of bacterial PHRs and consistent with distinct mechanisms for activities and regulation. Mutational analyses were combined with this prototypic structure for the (6-4) PHR/clock CRY cluster to identify structural and functional motifs: phosphate-binding and Pro-Lys-Leu protrusion motifs constricting access to the substrate-binding cavity above FAD, sulfur loop near the external end of the Trp electron-transfer pathway, and previously undefined C-terminal helix. Our results provide a detailed, unified framework for investigations of (6-4) PHRs and the mammalian CRYs. Conservation of key residues and motifs controlling FAD access and activities suggests that regulation of FAD redox properties and radical stability is essential not only for (6-4) photoproduct DNA repair, but also for circadian clock-regulating CRY functions. The structural and functional results reported here elucidate archetypal relationships within this flavoprotein family and suggest how PHRs and CRYs use local residue and cofactor tuning, rather than larger structural modifications, to achieve their diverse functions encompassing DNA repair, plant growth and development, and circadian clock regulation.     

In vitro reconstitution and crystal structure of p-aminobenzoate N-oxygenase (AurF) involved in aureothin biosynthesis

p-Aminobenzoate N-oxygenase (AurF) from Streptomyces thioluteus catalyzes the formation of unusual polyketide synthase starter unit p-nitrobenzoic acid (pNBA) from p-aminobenzoic acid (pABA) in the biosynthesis of antibiotic aureothin. AurF is a metalloenzyme, but its native enzymatic activity has not been demonstrated in vitro, and its catalytic mechanism is unclear. In addition, the nature of the cofactor remains a controversy. Here, we report the in vitro reconstitution of the AurF enzyme activity, the crystal structure of AurF in the oxidized state, and the cocrystal structure of AurF with its product pNBA. Our combined biochemical and structural analysis unequivocally indicates that AurF is a non-heme di-iron monooxygenase that catalyzes sequential oxidation of aminoarenes to nitroarenes via hydroxylamine and nitroso intermediates.     

Alpha-1-antitrypsin (AAT) and its stimulation in the liver of PiMZ phenotype individuals. A “recruitment-secretory block” (“R-SB”) phenomenon

ABSTRACT— PiMZ individuals in conditions of clinical stimulation show a peculiar immunohistochemical staining pattern for alpha-1-antitrypsin (AAT) in the liver: 1) the positivity involves large zones of parenchyma (up to 100% of hepatocytes); 2) in zone 2 and zone 3 hepatocytes the positivity appears in the form of crescents or rectilinear arrays along the sinusoids. This new pattern is designated type II, in contrast to type I which occurs in periportal hepatocytes in the form of inclusions spread over the whole cytoplasm. This peculiar staining pattern is associated with serum elevation of AAT and is considered to be an expression of liver reactivity. Type II positivity marks the hepatocytes which are newly recruited for the synthesis of AAT; owing to its defective export, the Z AAT is retained within the cell and detectable by immunohistochemistry. Thus this staining pattern is an expression of both recruitment for synthesis and block of secretion (“Recruitment-Secretory Block” “R-SB” phenomenon). In PiMZ individuals, the secretory block affects selectively and exclusively the Z fraction of AAT. At the EM level the vast majority of the retained protein is found in the RER, which represents the major and the earliest site of storage. Viewed in the framework of present knowledge of glycoprotein biosynthesis, the results of this study shed further light on the nature and cellular site of the Z AAT defect.     

黄海和渤海经济鱼类感染内弯宫脂线虫的调查

目的 调查我国黄海、渤海经济鱼类感染异尖科内弯宫脂线虫（Hysterothylacium aduncum）状况。方法 购买石岛渔港销售的鱼类93种941尾，分别解剖收集消化道内线虫，标本经乳酚酸透明，光镜下观察鉴定，同时用扫描电镜观察其超微结构。 结果 调查的93种鱼类中，有14种感染内弯宫脂线虫，占15.1%（14/93），其中感染率较高的为鮟鱇（Lophius litulon， 66.7%）、鳕鱼（Gadus macrocephalus， 47.5%）、马鲅（Scomberomorus niphonius，33.3%）等，首次发现内弯宫脂线虫可在软骨鱼史氏鳐（Raja smirnovi）体内寄生。此外不同宿主体内感染的内弯宫脂线虫形态特征有较大差异，如侧翼宽度以及肠盲囊的长度等有所不同。 结论 黄渤海经济鱼类感染内弯宫脂线虫的情况比较严重，不同宿主鱼类体内感染的内弯宫脂线虫可能是由不同虫体组成的复合体。     

Nonalcoholic Steatohepatitis (NASH) in OB/OB Mice Treated with Yo Jyo Hen Shi Ko (YHK): Effects on Peroxisome Proliferator-Activated Receptors (PPARs) and Microsomal Triglyceride Transfer Protein (MTP)

YHK has antioxidant properties, has a hypoglycemic effect, and may reduce plasma lipid levels. In this study, we examined the hepatic expression of PPAR-α and -γ and MTP in ob/ob mice receiving or not receiving YHK. Ob/ob mice were assigned to receive oral YHK (20 mg/kg/day) fed solution (methionine/choline-deficient [MCD] diet + YHK group) or vehicle (MCD group) by gavage for 4 weeks. Liver fragments were collected for histologic examination and mRNA isolation. PPAR-α and -γ and MTP gene expression was examined by RT-qPCR. YHK treatment was associated with NASH prevention, weight loss, and reduction of visceral fat and of serum concentrations of aminotransferases in comparison to the MCD group. YHK promoted an increment in PPAR-α and MTP and a decrement in PPAR-γ mRNA contents. These findings suggest that modulation of PPAR-α and -γ and MTP RNA expression may be implicated in the protective effect of YHK in experimental NASH, limiting hepatocyte lipid accumulation.     

Trends in Alcohol- and Drug-Related ED and Primary Care Visits: Data from Three U.S. National Surveys (1995–2005)

Objective: To evaluate trends in alcohol- and drug-related emergency department (ED) and primary care visits over the previous decade. Method: A trend analysis was conducted on substance-related health services visit data, based on self-reported drinking or drug use within six hours prior to an injury or illness event, from the 1995, 2000, and 2005 National Alcohol Surveys. Results: Although an upward trend was observed in alcohol-related ED visits from 1995 to 2005, this increase was not significant. A significant trend was found for drug-related ED visits from. 6% in 1995 to 3.7% in 2005 (p <. 01). In multiple logistic regression, year of survey (2000 vs. 1995) was positively predictive of drug-related ED visits, controlling for gender, age, ethnicity, and health insurance coverage; however, year of survey (2005 vs. 2000) was not significant. Conclusion: These data suggest that drug-related ED visits are continuing to increase, although the increase has not been as substantial between 2000 and 2005 as that which was observed between 1995 and 2000 and highlight the opportunity provided by the ED and primary care settings for screening and brief intervention for substance-related problems. These findings also suggest that Healthy People 2010 objectives calling for a reduction in substance-related visits may not be reached.     

Carfilzomib or bortezomib in combination with cyclophosphamide and dexamethasone followed by carfilzomib maintenance for patients with multiple myeloma after one prior therapy: results from a multicenter,phase II,randomized, controlled trial (MUKfive)

The proteasome inhibitors, carfilzomib and bortezomib, are widely used to treat myeloma but head-to-head comparisons have produced conflicting results. We compared the activity of these proteasome inhibitors in combination with cyclophosphamide and dexamethasone (KCd vs. VCd) in second-line treatment using fixed duration therapy and evaluated the efficacy of carfilzomib maintenance. MUKfive was a phase II controlled, parallel group trial that randomized patients (2:1) to KCd (n=201) or VCd (n=99); responding patients on carfilzomib were randomized to maintenance carfilzomib (n=69) or no further treatment (n=72). Primary endpoints were: (i) very good partial response (non-inferiority, odds ratio [OR] 0.8) at 24 weeks, and (ii) progression-free survival. More participants achieved a very good partial response or better with carfilzomib than with bortezomib (40.2% vs. 31.9%, OR=1.48, 90% confidence interval [CI]: 0.95, 2.31; non-inferior), with a trend for particular benefit in patients with adverse-risk disease. KCd was associated with higher overall response (partial response or better, 84.0% vs. 68.1%, OR=2.72, 90% CI: 1.62, 4.55, P=0.001). Neuropathy (grade ≥3 or ≥2 with pain) was more common with bortezomib (19.8% vs. 1.5%, P<0.0001), while grade ≥3 cardiac events and hypertension were only reported in the KCd arm (3.6% each). The median progression-free survival in the KCd arm was 11.7 months vs. 10.2 months in the VCd arm (hazard ratio [HR]=0.95, 80% CI: 0.77, 1.18). Carfilzomib maintenance was associated with longer progression-free survival, median 11.9 months vs. 5.6 months for no maintenance (HR 0.59, 80% CI: 0.46-0.77, P=0.0086). When used as fixed duration therapy in first relapase, KCd is at least as effective as VCd, and carfilzomib is an effective maintenance agent. This trial was registered with International Standard Randomised Controlled Trial Number (ISRCTN) identifier: ISRCTN17354232.     

Single nucleotide polymorphisms of matrix metalloproteinase 9 (MMP9) and tumor protein 73 (TP73) interact with Epstein-Barr virus in chronic lymphocytic leukemia: results from the European case-control study EpiLymph

Using EpiLymph case-control data, we found that chronic lymphocytic leukemia patients were more likely to have abnormal reactive serological patterns to Epstein Barr virus than controls. Here, we aimed to assess whether this association is modified by genetic variants. We examined 1,305 Single Nucleotide Polymorphisms from 300 selected genes related to various pathways in 240 cases and 513 controls from five European centers. In a recessive model, patients positive to aberrant antibody pattern and homozygous for rare genotypes in rs8113877T>G or rs17576A>G of the MMP9 gene were at highest risk of chronic lymphocytic leukemia. In a dominant model, TP73 showed the highest risk in patients positive to aberrant antibody pattern and homozygous for the wild-type genotype in rs1885859G>C or rs3765701A>T. All interactions were additive and no main effect was observed. The strong interactions observed may be indicative of a specific pathway in cancer genesis. Confirmation of these results is warranted.     

Comparative thyroid hormone concentration in maternal serum and yolk of the bonnethead shark (Sphyrna tiburo) from two sites along the coast of Florida

Maternally provisioned yolk hormones have been determined to play critical roles in development across vertebrate taxa. This study ascertained the presence and concentration of thyroid hormones triiodothyronine (T(3)) and thyroxine (T(4)) in the maternal serum and yolk of the developing placental viviparous shark Sphyrna tiburo from one site adjacent to Tampa Bay and another within Florida Bay, Florida, USA. The developmental profile of T(3) in yolk showed a steady increase from pre-ovulation to post-ovulation and peaked to its highest concentration during the pregnancy stage. There was an increase in the T(3)/T(4) ratio in yolk during the pregnancy stage which suggests a possible increase in the conversion of T(4) to T(3) within yolk, possible embryonic endogenous production, or passive uptake of T(3) from uterine fluids. Similar to the pattern seen in yolk, maternal serum T(3) concentrations tended to increase as development progressed. The concentration of T(3) and T(4) in yolk from Tampa Bay was consistently higher than in yolk from Florida Bay. The differences in the patterns of thyroid hormones from these two locations may explain previously reported differences in the rate of embryonic development in the two locations.     

Feasibility and toxicity of high-dose therapy (HDT) supported by peripheral blood stem cells in elderly patients with multiple myeloma and non-Hodgkin's lymphoma: survey from a single institution

The aim of this retrospective study was to investigate the feasibility of high-dose therapy (HDT) followed by peripheral blood stem cell transplantation (PBSCT) in elderly patients with hematological malignancies. From April 1998 to November 2001, 40 elderly patients (defined as > or =60 years) with non-Hodgkin's lymphoma (12 patients) and multiple myeloma (28 patients) were evaluated. Seven lymphoma and one myeloma patients were in complete remission (CR), 27 in partial remission (PR), two had stable disease (SD), and three progressive disease (PD). The median age was 65 years (range 60-71). Thirty-nine patients were mobilized with chemotherapy plus granulocyte-colony stimulating factor (G-CSF) and one with G-CSF alone. Patients received HDT including melphalan alone in 32 cases or combined with other drugs in six and BEAM in two. The median number of collected CD34(+) cells was 12.4 x 10(6)/kg (range 2.0-68.9). The median number of re-infused CD34(+) cells was 9.9 x 10(6)/kg (range 2.0-68.9). All patients engrafted after PBSC and the median time to neutrophil recovery (N > 500/micro l) and platelet recovery (PLT > 20,000/micro l) was 8 days (range 5-18) and 6 days (range 5-18), respectively. Nonhematological toxicity was mild and no patient died from transplant-related toxicity (TRM). Median duration of hospitalization was 18 days (range 12-24). To date, 32 patients are alive and eight died from disease progression at a median follow-up interval of 24 months. HDT supported by PBSC is a feasible procedure in selected elderly patients, and an age of more than 60 years should not be considered a contraindication for HDT.     

Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) DNA sequences are absent in leukapheresis products and ex vivo expanded CD34+ cells from multiple myeloma patients

Recently it was reported that Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) infects bone marrow (BM) dendritic cells (DC) in multiple myeloma (MM) patients and therefore might play a role in MM development. Because of the use of myeloid growth factors like GM-CSF and G-CSF for the mobilization of peripheral blood progenitor cells (PBPC), the subsequent increase of DC precursors might imply a risk for KSHV contamination in PBPC grafts. Therefore, in this study leukapheresis products and ex vivo cultured CD34+ cell suspensions were analysed. KSHV DNA could not be amplified in any of them.     

Interferon-γ+ and interleukin-2+ T cells in peripheral blood from multiple myeloma patients in relation to disease status and maintenance therapy with interferon-α2b (intron A)

Peripheral blood T cells from 83 patients with multiple myeloma (MM) were examined for the production of interferon-γ (INFγ) and interleukin-2 (IL-2) using three-colour flow cytometry. Comparisons were made between the percentage of cytokine-positive lymphocytes in normal donors and in patients during remission or relapse. Patients were divided into those who were on maintenance therapy with interferon-α2b (intron A) and those who had no further treatment after high-dose melphalan (HDM) with or without autologous bone marrow (ABMR) or peripheral blood stem cell rescue (PBSCR). The percentage of INFγ⁺/CD3⁺, INFγ⁺/CD45R0⁺/CD3⁺ and IL-2⁺/CD8⁺ was higher in patients on INFα2b during remission and relapse compared with normal donors (P < 0.005). During remission INFγ⁺/CD45R0⁺/CD3⁺ and IL-2⁺/CD8⁺ lymphocytes were higher in patients not on INFα2b (P < 0.05 and P < 0.005, respectively). In relapsed patients INFγ⁺/CD3⁺ and INFγ⁺/CD45R0⁺/CD3⁺ were increased in patients not taking INFα2b (P < 0.005). There was no significant difference between the percentages of cytokine-positive lymphocytes in patients taking or not taking INFα2b either during remission or relapse. Plasma IL-6 levels were similar in both groups of patients during remission. The data suggest that if maintenance therapy with INFα2b induces the synthesis of INFγ and IL-2 in vivo, the magnitude of the effect is small and may be unimportant in providing an anti-tumour effect in the majority of patients.     

Lipoprotein(a): Pathophysiology,measurement, indication and treatment in cardiovascular disease. A consensus statement from the Nouvelle Société Francophone d’Athérosclérose (NSFA)

Central Illustration. Pathophysiological pathways providing a causal link between high plasma concentrations of lipoprotein(a) (Lp(a)) and atherosclerotic vascular disease and aortic valve stenosis (AVS). Clinical outcomes are related to accelerated atherosclerosis complicated by atherothrombosis (myocardial infarction, stroke), peripheral artery disease (PAD) or aortic valve replacement (AVR) caused by valve calcification and aortic stenosis. Apo(a): apolipoprotein(a); LDL: low-density lipoprotein; OxPL: oxidized phospholipids; NSFA: Nouvelle Société Francophone d’Athérosclérose; SP: serine-protease domain; V: plasminogen kringle V (reproduced with permission).