orsai, the Drosophila homolog of human ETFRF1, links lipid catabolism to growth control.

BACKGROUND: Lipid homeostasis is an evolutionarily conserved process that is crucial for energy production, storage and consumption. Drosophila larvae feed continuously to achieve the roughly 200-fold increase in size and accumulate sufficient reserves to provide all energy and nutrients necessary for the development of the adult fly. The mechanisms controlling this metabolic program are poorly understood. RESULTS: Herein we identified a highly conserved gene, orsai (osi), as a key player in lipid metabolism in Drosophila. Lack of osi function in the larval fat body, the regulatory hub of lipid homeostasis, reduces lipid reserves and energy output, evidenced by decreased ATP production and increased ROS levels. Metabolic defects due to reduced Orsai (Osi) in time trigger defective food-seeking behavior and lethality. Further, we demonstrate that downregulation of Lipase 3, a fat body-specific lipase involved in lipid catabolism in response to starvation, rescues the reduced lipid droplet size associated with defective orsai. Finally, we show that osi-related phenotypes are rescued through the expression of its human ortholog ETFRF1/LYRm5, known to modulate the entry of ß-oxidation products into the electron transport chain; moreover, knocking down electron transport flavoproteins EtfQ0 and walrus/ETFA rescues osi-related phenotypes, further supporting this mode of action. CONCLUSIONS: These findings suggest that Osi may act in concert with the ETF complex to coordinate lipid homeostasis in the fat body in response to stage-specific demands, supporting cellular functions that in turn result in an adaptive behavioral response.

Plasmodium chabaudi-infected mice spleen response to synthesized silver nanoparticles from Indigofera oblongifolia extract.

Malaria is a worldwide serious-threatening infectious disease caused by Plasmodium and the parasite resistance to antimalarial drugs has confirmed a significant obstacle to novel therapeutic antimalarial drugs. In this article, we assessed the antioxidant and anti-inflammatory activity of nanoparticles prepared from Indigofera oblongifolia extract (AgNPs) against the infection with Plasmodium chabaudi caused in mice spleen. AgNPs could significantly suppress the parasitaemia caused by the parasite to approximately 98% on day 7 postinfection with P. chabaudi and could improve the histopathological induced spleen damage. Also, AgNPs were able to increase the capsule thickness of the infected mice spleen. In addition, the AgNPs functioned as an antioxidant agent that affects the change in glutathione, nitric oxide and catalase levels in the spleen. Moreover spleen IL1ß, IL-6 and TNF-α-mRNA expression was regulated by AgNPs administration to the infected mice. These results indicated the anti-oxidant and the anti-inflammatory protective role of AgNPs against P. chabaudi-induced spleen injury.

Predictive factors for mild forms of spondylotic cervical myelopathy treated conservatively or surgically.

A prospective 3-year randomized study comparing conservative and surgical treatment of spondylotic cervical myelopathy to establish predictive factors for outcome after conservative treatment and surgery. The clinical, electrophysiological and imaging parameters were examined to reveal how they characterized the clinical outcome. Statistically, pair-wise and multiple comparisons of different were used with the independent t-test and on one-way anova models followed by Tukey multiple-range tests. The patients with a good outcome in the conservatively treated group were of older age before treatment, had normal central motor conduction time (CMCT), and possessed a larger transverse area of the spinal cord. The patients with a good outcome in the surgically treated group had a more serious clinical picture (expressed in mJOA score and slower walk). Patients should rather be treated conservatively if they a spinal transverse area larger than 70 mm2, are of older age, and have normal CMCT. Surgery is more suitable for patients with clinically worse status and a lesser transverse area of spinal cord.

Predictive factors for spondylotic cervical myelopathy treated conservatively or surgically.

A prospective 3-year randomized study comparing conservative and surgical treatment of spondylotic cervical myelopathy to establish predictive factors for outcome after conservative treatment and surgery. The clinical, electrophysiological and imaging parameters were examined to reveal how they characterized the clinical outcome. The patients with a good outcome in the conservatively treated group were of older age before treatment, had normal central motor conduction time (CMCT), and possessed a larger transverse area of the spinal cord. The patients with a good outcome in the surgically treated group had a more serious clinical picture (expressed in mJOA score and slower walk). Patients should rather be treated conservatively if they have a spinal transverse area larger than 70 mm2, are of older age and have normal CMCT. Surgery is more suitable for patients with clinically worse status and a lesser transverse area of spinal cord.

Inhibition of liver carcinogenesis in Wistar rats by consumption of an aqueous extract from leaves of Ardisia compressa.

This study evaluates the chemopreventive effect of an aqueous extract of dried leaves of Ardisia compressa against liver cancer. A rat liver assay that mimics progressive forms of human disease was used as a carcinogenesis model. Forty-five male Wistar rats (180-200 g body weight) were injected intraperitoneally on day 1 with a single dose (100 mg/kg) of diethylnitrosamine (DEN), and also received via gavage 20 mg/kg acetylaminofluorene (2-AAF), on days 7, 8 and 9. The rats were randomly divided into four groups (n=15). Control groups (Group 1 and Group 2) had free access to water. Group 3 received 0.5% (w/v) of A. compressa tea for 10 days before treatment and during the study as the sole source of fluid until the rats were killed. A fourth group of 15 rats received no carcinogen or promoter but did receive 0.5%, (w/v) of A. compressa tea. All animals had 70% partial hepatectomy at day 10. The incidences of hepatocellular foci, nodules and carcinoma were significantly smaller in Group 3 than in Group 2 (P<0.01). A. compressa tea consumption alone (Group 4) did not induce the development of foci, nodules or carcinomas (P<0.01). The striking observation of this study was that consumption of A. compressa tea resulted in complete inhibition of the chemically-induced hepatocarcinogenesis in Wistar rats.

Prevalence of nutritional deficiencies in Mexican adolescent women with early and late prenatal care

The objective was to evaluate the prevalence of specific nutritional deficiencies in a group of pregnant adolescents according to the gestational age when they started to receive prenatal care. A group of 163 pregnant adolescents that attended the Instituto Nacional de Perinatología (Mexico City) for the first time to receive prenatal care was evaluated. An anthropometrical evaluation was performed and a blood sample taken to determine hemoglobin, ferritin, erythrocyte folate and plasma zinc to all cases. The mean age was 15 years (11 to 17 years). The mean gestational age when starting prenatal care was 27 +/- 7 gestation weeks and most of them tended to have low weight (97 +/- 12 per cent expected weight for height and gestational age). Eight of every ten adolescents had anemia and iron deficiency. Late prenatal care (> or = 25 weeks) was associated with the risk of presenting anemia OR 5.11 (CI 95 per cent 2.4-10.7) iron deficiency (OR 3.5; CI 95 per cent 1.7 to 7.1) and zinc deficiency (OR 2.9; CI 95 per cent 1.1 a 7.6). In relation to folate deficiency, the opposite effect was observed (OR 0.10; CI 95 per cent 0.02 a 0.48). Lack of opportune prenatal care was associated with the presence of iron and zinc depletion. Probably iron deficiency contributes to an erythrocyte folate accumulation.

Ontogeny constrains systemic protease inhibitor response in Nicotiana attenuata.

Protease inhibitors (PIs) are plant compounds that can inhibit proteases of mammal, insect, or pathogen origin and are frequently induced by mechanical wounding, insect feeding, or pathogen infection. Nicotiana attenuata is a species that induces nicotine, volatiles, and phenolics in response to damage. Here we examine the distribution of PIs in N. attenuata to determine if they are part of the induced response in this species and if this response is ontogenetically constrained. We found that N. attenuata shoot extracts inhibited trypsin (Tryp) and chymotrypsin (Chym) activities, while root extracts inhibited Tryp, Chym, and the bacterial protease subtilisin (Sub). The highest TrypPI levels were found at midday in the source-sink transition leaf, while older or younger leaves contained lower TrypPI levels and did not show significant diurnal fluctuations. Rosette plants, bolting plants, and flowering plants all contained TrypPIs in leaves, stems, and flowers, while seed capsules, seeds, and young seedlings did not contain any PIs. PIs in N. attenuata rosette plants were induced by Manduca sexta larval feeding, methyl jasmonate (MeJA) treatment, wounding, and application of M. sexta oral secretion and regurgitant. The response to MeJA application was stronger and longer lasting than to mechanical wounding. The direction and magnitude of the systemic response to mechanical wounding or larval damage depended on the age of the leaf that was damaged and the frequency of wounding. The systemic signal for TrypPI induction appears to follow source-sink relations in the plant and to be regulated by the octadecanoid pathway. Interestingly, by the time plants reach the flowering stage, they had lost the ability to increase PI levels after MeJA treatment. We concluded that plant ontogeny constrains both constitutive and inducible PI production in N. attenuata.

Arginine-based structures are specific inhibitors of cathepsin C. Application of peptide combinatorial libraries.

Novel synthetic peptide inhibitors of lysosomal cysteine proteinase cathepsin C have been designed through the use of soluble peptide combinatorial libraries. The uncovered structural inhibitory module consists of the N-terminal cluster of L-arginine residues. Its modification with D-amino acids or arginine derivatives did not increase the inhibition strength. Inhibitory potency of oligoarginines improves with the elongation of peptide chain reaching a maximum for octa-L-arginine. The oligoarginines specifically interact with the cathepsin C active site as shown by competitive-type inhibition kinetics (Ki approximately 10-5 M) and intrinsic fluorescence measurements. The inhibitory interaction of oligoarginines is established through the specific spatial contact of a net of guanidino groups in the arginine side-chains, as indicated by comparison with inhibitory action of low molecular mass guanidine derivatives (Ki approximately 10-3 M). Nonarginine polyionic compounds cannot mimic the inhibitory effect of oligoarginines. The arginine-based peptide inhibitors were selective towards cathepsin C among other cysteine proteinases tested.

Report on a 3-year follow-up zidovudine (AZT) treatment in a group of HIV-positive patients with congenital clotting disorders.

Eleven (50%) of 22 HIV-seropositive patients suffering from congenital coagulation defects and followed at the Hemophilia Center of Padua met the eligibility criteria for zidovudine (AZT) therapy. A 3-year clinical and laboratory follow up is described. Mean length of AZT treatment was 14.3 +/- 11 months. Three patients were enrolled at the latest stages (CDC stage IV) of HIV disease. They showed no clinical improvement after AZT administration and died with AIDS. One CDC stage III patient died from a high-grade non-Hodgkin's lymphoma (NHL) which suddenly developed 3 months after starting AZT. Seven patients began antiretroviral treatment when they were mild or asymptomatic for HIV infection (CDC stage II and III). None developed any sign of HIV disease progression on the basis of CDC criteria. Moreover, AZT administration induced an improvement of the humoral markers related to HIV disease, as CD4 T-lymphocyte count, serum beta 2-microglobulin (B2M) and, although only for few months, neopterin (Np) levels. A mild thrombocytopenia due to HIV infection was detected in 5 patients. In all cases AZT treatment was effective in increasing and/or normalizing the platelet count. Reduced daily dose AZT (600-500 mg/day) appeared to be well tolerated and of minimal toxicity as compared to the higher dose (1200 mg/day). In our study, the zidovudine-induced bone marrow suppression, namely severe anemia or pancytopenia, was the major side-effect limiting tolerance of the higher dose AZT.

Bleeding as a cause of death in HIV-seropositive patients with congenital clotting disorders.

The causes of death in a group of HIV-seropositive patients suffering from congenital clotting disorders (cCD) were studied. During the past 6 years, we have followed 19 patients with cCD and HIV infection. Eight patients fulfilled revised CDC criteria for AIDS, 6 subjects reached stage III of CDC, and 5 remained asymptomatic (CDC stage II). All patients who developed AIDS died. In 5 patients, the terminal cause of death was a severe haemorrhage (hematemesis, melena or haemoptysis) after gastrointestinal or lung opportunistic infections. Two other patients died as a consequence of disseminated infections but without significant bleeding. Only one subject died due to neoplastic disease. In the first stages of CDC (II and III), no increase in bleeding symptoms were seen in cCD HIV infected patients. The risk of haemorrhages is, however, increased in AIDS patients. Adequate replacement therapy should be started early whenever severe opportunistic infections appear.

New trypsin inhibitors are present in the coelomic fluid of the earthworm Lumbricus terrestris.

Proteinase-inhibiting components of the coelomic fluid of the earthworm Lumbricus terrestris were examined. Inhibition of proteinases of serine, aspartate and thiol families was tested. Very strong inhibition was observed only in the case of trypsin. Additional data suggest that the inhibition is related to proteins of molar mass of 42 kDa and 20 kDa, respectively. These two proteins are present in the coelomic fluid in several forms which differ in their isoelectric points.

S-S bridges of cathepsin B and H from bovine spleen: a basis for cathepsin B model building and possible functional implications for discrimination between exo- and endopeptidase activities among cathepsins B, H and L.

Bovine spleen cathepsin B contains 7 disulfide bridges. Using different chemical and enzymatic cleavage methods we isolated fragments representing the individual disulfides: Cys14-Cys43, Cys26-Cys71, Cys62-Cys128, Cys63-Cys67, Cys100-Cys132, Cys108-Cys119, and Cys148-Cys252. A similar line of approach was applied to determine the S-S bridges of bovine spleen cathepsin H: Cys23-Cys66, Cys57-Cys99, Cys157-Cys207, and Cys212-Cys5A, where Cys5A is located in the propart portion of the procathepsin H chain. On the basis of the knowledge of the S-S bridges of cathepsin B a novel sequence alignment of papain and cathepsin B has been proposed. This enabled us to construct a reasonable 3D-model of cathepsin B and propose the region (a 18 residue insertion between Glu89 and Gly90 of papain) responsible for the carboxypeptidase activity of cathepsin B functioning as a "closure". A similar approach was applied to explain the aminopeptidase activity of cathepsin H. A general model of steric regulation of accessibility of the preformed "endopeptidase-like" binding cleft by distant parts of the polypeptide chain of the proteinases discussed is proposed as a factor determining the mode of binding and thus cleavage of polypeptide substrates.

Effect of buflomedil, dipyridamole and indobufene on hemorrheologic and blood coagulation parameters in patients who underwent vascular surgery for severe atherosclerosis.

We studied 30 patients (age range 58 to 64 years) who underwent vascular surgery for peripheral vascular disease (24 patients) and for carotid stenosis (6 patients). At the time of the study, all subjects have been taking antiplatelet agents for at least three months. They were divided into three groups of ten patients each: the first group received buflomedil (BUF), the second dipyridamole (DIP) and the third indobufene (IND), as treatment before vascular surgery. We investigated the influence of these drugs on hemorrheologic (hematocrit, whole blood and plasma viscosity) and blood coagulation (partial thromboplastin time, prothrombin time and bleeding time) parameters in all patients. In our experience, none of the three drugs influenced any tested parameters, which were within normal limits: among them we noted only a statistically significant difference in bleeding time between patients treated with BUF and IND.

Portal vein thrombosis in a patient with severe haemophilia A and post-hepatitis liver cirrhosis.

The occurrence of a portal vein thrombosis in a haemophilia A patient is reported. The patient, a 53 year old male, had been followed by us for the past 20 years in our out-patient Clinic. He was hospitalized recently for a suspected hepatic cirrhosis. Severe ascites, hepato-splenomegaly together with weight loss and mild fever were present. During the hospitalization, an ultrasound and CT scan of the liver confirmed the cirrhotic pattern and showed the presence of a portal vein thrombosis. There were no changes in the underlying coagulation defect, in fact, the patient had recurrent haemarthrosis. Furthermore, with the ultrasound examination, some focal hepatic lesions--probably due to a hepatocellular carcinoma--were also observed. The patient died because of massive haematemesis due to rupture of oesophageal varices.

Variability of plasma viscosity in monoclonal IgM gammopathies.

Hyperviscosity syndromes can caused by both plasmatic and cellular factors. We have studied 20 patients affected by IgM gammopathy of different origin and 12 healthy subjects matched for sex and age, in order to evaluate the relation between paraprotein levels and plasma viscosity. We have observed a significant plasma viscosity increase only in 14 patients with monoclonal IgMk gammopathy. In the same patients was also evident an hyperviscosity syndrome. In the other 6 patients, with monoclonal IgM or polyclonal gammopathy and without clinical symptoms, plasma viscosity was only slightly increased. We have also observed a significant correlation between IgM and light chains (kappa, lambda) serum level and increased plasma viscosity. These results suggest that one can't consider all IgM gammopathies as cause of hyperviscosity syndrome.

Rhinophyma-like cryptococcal infection as an early manifestation of AIDS in a hemophilia B patient.

A hemophilia B patient, seropositive for HIV antibodies since 1984, came to us in March 1989 with a severe necrotizing lesion of the nose. It was an erythematous lesion and looked like rhinophyma. Microbiological examination of the skin biopsy showed the presence of Cryptococcus neoformans. At the time of the study, the patient was in partial remission after 2 weeks of therapy with fluconazole per os 400 mg/day. He will be treated with the same therapy at maintenance dose (200 mg/day) for a long period.

Primary structure of cathepsin D inhibitor from potatoes and its structure relationship to soybean trypsin inhibitor family.

A novel effective procedure for the purification of cathepsin D inhibitor from potatoes (PDI) was developed. The amino acid sequence of PDI was determined by analysis of the cyanogen bromide digest and of the limited tryptic and chymotryptic digest of the protein. The inhibitor is a single polypeptide chain protein consisting of 188 residues with a simple sugar moiety attached to Asn-19. The tentative disulfide pairings are also suggested. The sequence data clearly indicate that PDI is homologous with the soybean trypsin inhibitor (STI) (Kunitz) family. The active center of PDI for trypsin inhibition was identified as Pro-Val-Arg-Phe in analogy to STI.

Iron stores in remunerated blood donors as evaluated by plasma ferritin levels.

This study was undertaken to investigate body iron stores in so-called remunerated blood donors as well as to evaluate the sensitivity of hemoglobin determination in detecting iron deficiency in two populations of blood donors. The authors studied 522 male donors who were divided into three groups: Group I, first-time volunteer donors with hemoglobin levels greater than or equal to 13 g per dl; Group II, remunerated donors with hemoglobin levels greater than or equal to 13 g per dl; and Group III, remunerated donors rejected because their hemoglobin levels were less than 13 g per dl. Iron stores were evaluated with an enzyme-linked immunosorbent assay for plasma ferritin. In Group I, 4.5 percent were iron-deficient with a mean ferritin value of 55.3 ng per ml; in Group II, 59.7 percent were iron deficient with a mean ferritin level of 17.4 ng per ml, and in Group III, 82.5 percent were iron-deficient and the mean ferritin level was 8.4 ng per ml. The last values represent the highest percentage of iron deficiency and the lowest mean ferritin value thus far reported. In Group I, hemoglobin determination had a sensitivity of 95 percent in detecting iron deficiency, but in Group II had only 40 percent sensitivity. These results indicate that a more accurate and reliable test, such as a plasma or serum ferritin determination, may be necessary to detect iron deficiency in blood donors when they donate more than five times per year, particularly those who are remunerated.