ABSTRACT
OBJECTIVE: Treacher Collins syndrome (TCS) is a rare congenital disorder of craniofacial development. TCS occurs with an incidence of 1:50,000, and more than 60% of TCS cases have no previous family history and arise as the result of de novo mutations. The high rate of de novo mutations, together with the extreme variability in the degree to which individuals can be affected, makes the provision of genetic counseling extremely complicated. Consequently, every case of TCS is unique and needs to be assessed individually. Patients with TCS frequently undergo multiple reconstructive surgeries from birth through adulthood, which rarely are fully corrective in the long-term. The nascent field of regenerative medicine offers the promise to improve some of these treatments. In particular, structural fat grafting (SFG) seems to be a good strategy not only to restore the normal volume and contour of the face, but also to provide a source of adipose-derived stem cells (ADSCs) with a multilineage differentiation potential. In this work, we present genetical analyses of ADSC affected by TCS. MATERIALS AND METHODS: ADSCs from were analyzed for their stemness properties and shared many characteristics with those of a healthy subject. Screening of the genome of the TCS patient using array-Comparative Genomic Hybridization allowed us to identify some chromosomal imbalances that are probably associated with TCS. RESULTS: We found that some alterations, involving the TIMELESS gene, were usually associated with embryonic stem cells. CONCLUSIONS: With the aim to improve the final results, we need to consider combining knowledge of genetic alterations and expression profiles as a fundamental step before starting with surgical procedures.
Subject(s)
Mandibulofacial Dysostosis , Plastic Surgery Procedures , Female , Humans , Mandibulofacial Dysostosis/etiology , Mandibulofacial Dysostosis/genetics , Comparative Genomic Hybridization , Mutation , Stem CellsABSTRACT
OBJECTIVE: A hernia of the abdominal wall is an opening of the muscles in the abdominal wall, which is frequently treated via the application of a surgical mesh. The purpose of this research is to study how human adipose-derived stem cells (hADSCs) interact with Phasix™ Mesh, a commercially available mesh for hernia repair. Studying how cells derived from the abdominal region behave with Phasix™ Mesh is crucial to improve the state of the art of current surgery and achieve effective tissue restoration. MATERIALS AND METHODS: hADSCs were seeded onto Phasix™ Mesh, a fully resorbable surgical mesh of poly (4-hydroxybutyric acid) (P4HB). Cell viability was assessed through MTT assay, and cell growth and adhesion were evaluated via multiple imaging techniques and gene imaging profiling. RESULTS: Results confirm that the nets support cells proliferation, extracellular matrix production and increasing of angiogenetic factor. CONCLUSIONS: Butyric acid-based nets are promising scaffolds for abdominal wall reconstruction.
Subject(s)
Abdominal Wall , Hernia, Ventral , Humans , Abdominal Wall/surgery , Tissue Engineering , Butyric Acid , Herniorrhaphy/methods , Prostheses and Implants , Surgical Mesh , Hernia, Ventral/surgeryABSTRACT
Twenty-four lambs (Ovis aries) were used in a 45-day finishing study to evaluate the effects of feeding diets high in linoleic acid (C(18:2), omega-6) on liver lipid composition and on lipogenic enzyme activities in subcellular fractions of liver. Lambs were fed either a 5% safflower oil (SO, high linoleic acid) supplemented diet or a control diet without added oil. SO feeding caused a reduction in the amount of serum and liver triacylglycerols and cholesterol, whereas the level of phospholipids in both tissues was hardly affected. In liver of SO-treated lambs an increase in the levels of C(18:2) and arachidonic acid (C(20:4), omega-6), together with a simultaneous decrease of saturated fatty acids, was observed. In comparison to rat liver, rather low activities of enzymes in the pathway for de novo fatty acid synthesis, i.e. acetyl-CoA carboxylase and fatty acid synthase, were found in lamb-liver cytosol. Both enzyme activities, as well as those of the NADPH-furnishing enzymes, were significantly reduced by SO feeding. In contrast, microsomal and especially mitochondrial fatty acid chain elongation activity, the latter being much higher than that of rat liver, were significantly increased in SO-treated lambs. In these animals, a stimulation of triangle up(9)-desaturase activity was observed in liver microsomes.
Subject(s)
Acetyl-CoA Carboxylase/metabolism , Dietary Fats/pharmacology , Fatty Acid Synthases/metabolism , Fatty Acids, Omega-6/pharmacology , Lipid Metabolism/drug effects , Liver/enzymology , Sheep, Domestic/metabolism , Stearoyl-CoA Desaturase/metabolism , Animals , Diet , Male , RatsABSTRACT
We studied the involvement of oxidative stress in chronic idiopathic urticaria (CIU), assessing the activities of superoxide dismutase (SOD) and glutathione and the levels of malondialdeyde (MDA), a marker of lipid peroxidation, in samples taken from lesional skin (n = 16) and nonlesional skin (n = 11) of CIU patients. The activity of SOD and glutathione and the levels of MDA were markedly increased in lesional skin as compared with skin of healthy subjects, whereas no differences were detected between nonlesional skin of CIU patients and control samples. Immuno-dot blot assay revealed an up-regulation of Mn-SOD expression in lesional skin. These findings show that oxidative stress is crucially involved in CIU. The evidence of lipid peroxidation and compensatory increase of Mn-SOD and glutathione activities in lesional skin, in the absence of any alteration in uninvolved skin, suggests that oxidative stress is secondary to the development of inflammation.
Subject(s)
Oxidative Stress , Superoxide Dismutase/metabolism , Urticaria/enzymology , Adult , Biomarkers/analysis , Chronic Disease , Female , Glutathione/metabolism , Humans , Lipid Peroxidation , Male , Malondialdehyde/metabolism , Middle Aged , Skin/metabolism , Urticaria/physiopathologyABSTRACT
The mitochondrial F(1)F(o) ATP synthase complex has a key role in cellular energy metabolism. The general architecture of the enzyme is conserved among species and consists of a globular catalytic moiety F(1), protruding out of the inner side of the membrane, a membrane integral proton translocating moiety F(o), and a stalk connecting F(1) to F(o). The X-ray crystallographic analysis of the structure of the bovine mitochondrial F(1) ATPase has provided a structural basis for the binding-change rotary mechanism of the catalytic process in F(1), in which the gamma subunit rotates in the central cavity of the F(1) alpha3/beta3 hexamer. Rotation of gamma and eta subunits in the E. coli enzyme and of, gamma and delta subunits in the mitochondrial enzyme, is driven, during ATP synthesis, by proton motive rotation of an oligomer of c subunits (10-12 copies) within the F(o) base piece. Average analysis of electron microscopy images and cross-linking results have revealed that, in addition to a central stalk, contributed by gamma and delta/eta subunits, there is a second lateral one connecting the peripheries of F(o) and F(1). To gain deeper insight into the mechanism of coupling between proton translocation and catalytic activity (ATP synthesis and hydrolysis), studies have been undertaken on the role of F(1) and F(o) subunits which contribute to the structural and functional connection between the catalytic sector F(1) and the proton translocating moiety F(o). These studies, which employed limited proteolysis, chemical cross-linking and functional analysis of the native and reconstituted F(1)F(o) complex, as well as isolated F(1), have shown that the N-terminus of alpha subunits, located at the top of the F(1) hexamer is essential for energy coupling in the F(1)F(o) complex. The alpha N-terminus domain appears to be connected to F(o) by OSCP (F(o) subunit conferring sensitivity of the complex to oligomycin). In turn, OSCP contacts F(o)I-PVP(b) and d subunits, with which it constitutes a structure surrounding the central gamma and delta rotary shaft. Cross-linking of F(o)I-PVP(b) and gamma subunits causes a dramatic enhancement of downhill proton translocation decoupled from ATP synthesis but is without effect on ATP driven uphill proton transport. This would indicate the existence of different rate-limiting steps in the two directions of proton translocation through F(o). In mitochondria, futile ATP hydrolysis by the F(1)F(o) complex is inhibited by the ATPase inhibitor protein (IF(1)), which reversibly binds at one side of the F(1)F(o) connection. The trans-membrane deltapH component of the respiratory deltap displaces IF(1) from the complex; in particular the matrix pH is the critical factor for IF(1)association and its related inhibitory activity. The 42L-58K segment of the IF(1) has been shown to be the most active segment of the protein; it interacts with the surface of one alpha/beta pairs of F(1), thus inhibiting, with the same pH dependence as the natural IF(1), the conformational interconversions of the catalytic sites involved in ATP hydrolysis. IF(1) has a relevant physiopathological role for the conservation of the cellular ATP pool in ischemic tissues. Under these conditions IF(1), which appears to be over expressed, prevents dissipation of the glycolytic ATP.
Subject(s)
Mitochondrial Proton-Translocating ATPases/chemistry , Mitochondrial Proton-Translocating ATPases/metabolism , Proton-Motive Force , Amino Acid Sequence , Animals , Models, Molecular , Molecular Sequence Data , Protein Binding , Protein Structure, Quaternary , Protein Subunits , Proteins/metabolism , ATPase Inhibitory ProteinABSTRACT
A study is presented of the activity and temperature dependence of the ATPase inhibitor protein (IF(1)) from bovine heart mitochondria and of synthetic partial IF(1) peptides. The results show that the IF(1)-(42-58) peptide is the most potent inhibitory domain of IF(1).
Subject(s)
Adenosine Triphosphatases/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Mitochondria, Heart/metabolism , Peptide Fragments/chemistry , Proteins/chemistry , Proteins/metabolism , Amino Acid Sequence , Animals , Cattle , Enzyme Inhibitors/isolation & purification , Intracellular Membranes/chemistry , Intracellular Membranes/metabolism , Kinetics , Mitochondria, Heart/chemistry , Molecular Sequence Data , Peptide Fragments/pharmacology , Proteins/isolation & purification , Thermodynamics , ATPase Inhibitory ProteinABSTRACT
We have studied the functional effect of limited proteolysis by trypsin of the constituent subunits in the native and reconstituted F1F0 complex and isolated F1 of the bovine heart mitochondrial ATP synthase (EC 3.6.1.34). Chemical cross-linking of oligomycin-sensitivity conferring protein (OSCP) with other subunits of the ATP synthase and the consequent functional effects were also investigated. The results obtained show that the alpha subunit N-terminus is essential for the correct, functional connection of F1 to F0. The alpha-subunit N-terminus contacts OSCP which, in turn, contacts the F0I-PVP(b) and the F0-d subunits. The N-terminus of subunit alpha, OSCP, a segment of subunit d and the C-terminal and central region of F0I-PVP(b) subunits are peripherally located with respect to subunits gamma and delta which are completely shielded in the F1F0 complex against trypsin digestion. This qualifies the N-terminus of subunit alpha, OSCP, subunit d and F0I-PVP(b) as components of the lateral element of the stalk. These subunits, rather than being confined at one side of the complex which would leave most of the central part of the gamma subunit uncovered, surround the gamma and the delta subunits located in the central stalk.
Subject(s)
Carrier Proteins , Mitochondria/enzymology , Proton-Translocating ATPases/chemistry , Adenosine Triphosphatases/physiology , Animals , Cattle , Cross-Linking Reagents/pharmacology , Dose-Response Relationship, Drug , Electrophoresis, Polyacrylamide Gel , Enzyme Inhibitors/pharmacology , Immunoblotting , Intracellular Membranes/enzymology , Kinetics , Light , Liposomes/drug effects , Membrane Proteins/physiology , Mitochondrial Proton-Translocating ATPases , Oligomycins/pharmacology , Protein Structure, Tertiary , Proton-Translocating ATPases/physiology , Protons , Time Factors , Trypsin/pharmacologyABSTRACT
In hypothyroid rats, partial hepatectomy does not induce liver regeneration until 120 h after surgical operation. when, instead, in normal rats a complete recovery of the liver mass, in this interval, is observed. In normal rats, a good efficiency of mitochondrial oxidative phosphorylation is needed as an energy source for liver regeneration (Guerrieri, F. et al., 1995); in hypothyroid rats the efficiency of mitochondrial oxidative phosphorylation is low in the 0-120 h interval after partial hepatectomy. This low efficiency of oxidative phosphorylation appears to be related to a low mitochondrial content of F0F1-ATP synthase, in liver of hypothyroid rats, which does not recover after partial hepatectomy. In the liver of hypothyroid rats, low levels of the nuclear-encoded mitochondrial catalytic betaF1 subunit and of its transcript are observed and they do not increase, as occurs in normal rats, after partial hepatectomy.
Subject(s)
Gene Expression , Hypothyroidism/enzymology , Liver Regeneration/physiology , Mitochondria, Liver/enzymology , Proton-Translocating ATPases/genetics , Adenosine Triphosphate/metabolism , Animals , Antithyroid Agents/adverse effects , Disease Models, Animal , Hepatectomy , Hypothyroidism/chemically induced , Liver/enzymology , Liver/physiology , Male , Propylthiouracil/adverse effects , Rats , Rats, Wistar , Time FactorsABSTRACT
The structural and functional connection between the peripheral catalytic F1 sector and the proton-translocating membrane sector F0 of the mitochondrial ATP synthase is reviewed. The observations examined show that the N-terminus of subunit gamma, the carboxy-terminal and central region of F0I-PVP(b), OSCP, and part of subunit d constitute a continuous structure, the lateral stalk, which connects the peripheries of F1 to F0 and surrounds the central element of the stalk, constituted by subunits gamma and delta. The ATPase inhibitor protein (IF1) binds at one side of the F1F0 connection. The carboxy-terminal segment of IF1 apparently binds to OSCP. The 42L-58K segment of IF1, which is per se the most active domain of the protein, binds at the surface of one of the three alpha/beta pairs of F1, thus preventing the cyclic interconversion of the catalytic sites required for ATP hydrolysis.
Subject(s)
Mitochondria/enzymology , Proton-Translocating ATPases/chemistry , Proton-Translocating ATPases/metabolism , Animals , Catalytic Domain , Crystallography, X-Ray , Escherichia coli/enzymology , Kinetics , Models, Molecular , Oligomycins/pharmacology , Peptide Fragments/chemistry , Protein SubunitsABSTRACT
A study is presented on the effect of diamide-induced disulfide cross-linking of F(1)-gamma and F(0)I-PVP(b) subunits on proton translocation in the mitochondrial ATP synthase. The results show that, upon cross-linking of these subunits, whilst proton translocation from the A side to the B F(1) side is markedly accelerated with decoupling of oxidative phosphorylation, proton translocation in the reverse direction, driven by either ATP hydrolysis or a diffusion potential, is unaffected. These observations reveal further peculiarities of the mechanism of energy transfer in the ATP synthase of coupling membranes.
Subject(s)
Disulfides/chemistry , Mitochondria/enzymology , Proton-Translocating ATPases/chemistry , Proton-Translocating ATPases/metabolism , Protons , Animals , Biological Transport/drug effects , Cattle , Cross-Linking Reagents/pharmacology , Diamide/pharmacology , Kinetics , Oligomycins/pharmacology , Sulfhydryl Reagents/metabolism , Uncoupling Agents/pharmacologyABSTRACT
Diamide treatment of the F0F1-ATP synthase in "inside out" submitochondrial particles (ESMP) in the absence of a respiratory Delta mu H+ as well as of isolated Fo reconstituted with F1 or F1-gamma subunit results in direct disulfide cross-linking between cysteine 197 in the carboxy-terminal region of the F0I-PVP(b) subunit and cysteine 91 at the carboxyl end of a small alpha-helix of subunit F1-gamma, both located in the stalk. The F0I-PVP(b) and F1-gamma cross-linking cause dramatic enhancement of oligomycin-sensitive decay of Delta mu H+. In ESMP and MgATP particles the cross-linking is accompanied by decoupling of respiratory ATP synthesis. These effects are consistent with the view that F0I-PVP(b) and F1-gamma are components of the stator and rotor of the proposed rotary motor, respectively. The fact that the carboxy-terminal region of F0I-PVP(b) and the short alpha-helix of F1-gamma can form a direct disulfide bridge shows that these two protein domains are, at least in the resting state of the enzyme, in direct contact. In isolated F0, diamide also induces cross-linking of OSCP with another subunit of F0, but this has no significant effect on proton conduction. When ESMP are treated with diamide in the presence of Delta mu H+ generated by respiration, neither cross-linking between F0I-PVP(b) and F1-gamma subunits nor the associated effects on proton conduction and ATP synthesis is observed. Cross-linking is restored in respiring ESMP by Delta mu H+ collapsing agents as well as by DCCD or oligomycin. These observations indicate that the torque generated by Delta mu H+ decay through Fo induces a relative motion and/or a separation of the F0I-PVP(b) subunit and F1-gamma which places the single cysteine residues, present in each of the two subunits, at a distance at which they cannot be engaged in disulfide bridging.
Subject(s)
Mitochondria, Heart/enzymology , Proton-Translocating ATPases/chemistry , Proton-Translocating ATPases/metabolism , Adenosine Triphosphate/biosynthesis , Animals , Cattle , Cross-Linking Reagents/metabolism , Diamide/pharmacology , Disulfides/chemistry , Electron Transport/drug effects , Mitochondria, Heart/drug effects , Proton Pumps/chemistry , Proton Pumps/metabolism , Submitochondrial Particles/drug effects , Submitochondrial Particles/enzymology , Sulfhydryl Reagents/metabolismABSTRACT
Capillary electrophoresis (CE) has emerged as an important technique applied to analytical chemistry and quality control. In the cosmetic field ascorbic acid is important for soothing, bleaching and for its scavenger activity. Ascorbic acid is responsible for great number of physiological oxidations in electron transfer reactions. Many natural substances have a functional activity which is very difficult to preserve or even to prolong with the requested and expected treatment. This paper reports the study of capillary electrophoresis for the characterization of ascorbic acid as the active ingredient in a new delivery system that is able to preserve stability and to prolong release of ascorbic acid according to the concentration needs of a cosmetic formulation.
ABSTRACT
In our work a non-classical experimental design was applied to obtain lipsticks endowed with particular characteristics. Our aim was to formulate lipsticks that leave a brilliant and shiny colour application and have a transparent look. The emollient substances and the waxes (consistency factors) were identified as the main variables of the system. A two phase experimental strategy was thought out: the optimal quantities of consistency factors were selected using a Doehlert experimental matrix, whereas the correct mixtures of emollients were determined using a Scheffé simplex-centroid design. These two design were combined and a set of 49 experiments was obtained. The experiments carried out allowed the definition of a zone of two phases in which the objectives were attained: the correct types and appropriate quantities of emollients and waxes were determined. To find a possible correlation between some mixtures and the lipsticks' sensorial behaviour, differential scanning calorimetry was used. These results, in addition to those obtained using the experimental design allowed us to select the best lipstick formula. (c) Rapid Science Ltd. 1998.
ABSTRACT
BACKGROUND AND AIM: To evaluate the efficacy of premedication with midazolam (mdz) administered using a nasal route compared to diazepam (dz) administered by mouth in children of different ages. EXPERIMENTAL DESIGN: A comparative type study was performed in randomly selected pediatric patients undergoing surgery. The study lasted 3 months. SETTING: Recovery room and operating theatre for Pediatric Surgery and ENT. PATIENTS: A total of 248 patients were studied, divided into 3 age groups: group A were aged under 2 years; group B were pre-school age and group C were school-age. OPERATIONS: Two subgroups were formed based on the premedication used: group M = 0.2 mg/kg of mdz using a nasal route on arrival in the operating unit; group D = 0.2 mg/kg of dz per os 45' before induction. PARAMETERS STUDIED: In addition to acceptance of treatment, which was deemed to be good, poor or refused, the authors evaluated the level of sedation (score from 5 to 1: awake-asleep), anxiety on entering SO (score from 1 to 4: none-excessive) and the level of collaboration during the induction of general anesthesia (score 1-4: excellent-nil). RESULTS: The nasal route was well accepted by 59% of patients in group A, 62% of group B and 97% of group C. Statistical analysis using Kruskall Wallis test showed significant differences in groups A and B between the two subgroups M and D for all the parameters studied, whereas there were no significant differences in group C. CONCLUSIONS: Premedication with mdz using a nasal route was safe and efficacious, above all in early and later infancy.
Subject(s)
Adjuvants, Anesthesia/administration & dosage , Midazolam/administration & dosage , Preanesthetic Medication , Administration, Intranasal , Aging , Child , Child, Preschool , Humans , InfantABSTRACT
Peptide segments of the inhibitor protein (IF1) of the F0F1 ATP synthase complex from bovine-heart mitochondria have been constructed by chemical synthesis. The IF1-(42-58)-peptide was equally effective as IF1 in inhibiting the ATPase activity of both the F0F1 complex in the mitochondrial membrane deprived of IF1 (SMP) and soluble F1. The IF1-(22-46)-peptide inhibited the ATPase activity in the soluble F1 but had no effect on either the ATPase activity or H+ conduction in SMP. Substitution of the His or Lys residues with Ala in the IF1-(42-58)-peptide decreased the inhibition of ATP hydrolysis. The inhibition exerted by the IF1-(42-58)-peptide on ATP hydrolysis in SMP exhibited a pH dependence, similar to that observed with IF1, which was lost upon replacement of His or Lys with Ala. In soluble F1, inhibition of ATP hydrolysis by IF1, the IF1-(42-58)-peptide and the IF1-(22-46)-peptide was pH dependent when F1 was first incubated with ATP. The IF1-(42-58)-peptide also caused inhibition of passive H+ conduction in SMP. This activity of the synthetic peptide was weaker, as compared to that of IF1, and practically unaffected by substitution of His or Lys with Ala. An antibody against the IF1-(42-58)-synthetic peptide stimulated ATP hydrolysis in the membrane-bound F0F1 complex with associated IF1 but was without effect on H+ conduction. An antibody against IF1 stimulated both processes.
Subject(s)
Enzyme Inhibitors/chemistry , Proteins/chemistry , Proton-Translocating ATPases/antagonists & inhibitors , Adenosine Triphosphate/metabolism , Amino Acid Sequence , Animals , Antibodies/immunology , Cattle , Enzyme Inhibitors/metabolism , Enzyme Inhibitors/pharmacology , Hydrogen-Ion Concentration , Mitochondria, Heart/enzymology , Molecular Sequence Data , Mutation , Peptide Fragments/chemical synthesis , Peptide Fragments/chemistry , Peptide Fragments/metabolism , Peptide Fragments/pharmacology , Proteins/metabolism , Proteins/pharmacology , Proton-Translocating ATPases/metabolism , Protons , ATPase Inhibitory ProteinABSTRACT
The DCCD-sensitive proton permeability of chromatophores, from a green strain of Rhodobacter Capsulatus is potentiometrically detected following the proton release induced by a transmembrane diffusion potential imposed by a valinomycin-mediated potassium influx with a procedure already used for bovine heart submitochondrial particles (ESMP) and vesicles from Escherichia coli (Zanotti et al. (1994) Eur. J. Biochem. 222, 733-741). In the photosynthetic system, addition of increasing amounts of DCCD inhibits, with a similar titre, both proton permeability and MgATP-dependent ATPase activity as detected in the dark. The titre for 50% inhibition coincides with that obtained measuring proton permeability and ATP hydrolysis in ESMP. Upon removal of F1, the passive proton permeability is much less sensitive to DCCD in chromatophores than in USMP, suggesting that in chromatophores the F1-Fo interaction shapes the DCCD-sensitive proton conducting pathway. Addition of the purified mitochondrial FoI-PVP and oligomycin sensitivity-conferring (OSCP) proteins to the F1 stripped chromatophores restored the sensitivity of proton permeability to DCCD detected in untreated chromatophores. Analysis of the binding of 14C[DCCD] on F1 stripped chromatophores shows that the increase of DCCD sensitivity of proton permeability, caused by addition of mitochondrial Fo proteins, is related to an increase of the binding of the inhibitor to subunit c of Fo sector of ATP synthase complex.
Subject(s)
Bacterial Chromatophores/drug effects , Bacterial Chromatophores/metabolism , Dicyclohexylcarbodiimide/pharmacology , Proton-Translocating ATPases/metabolism , Rhodobacter capsulatus/drug effects , Rhodobacter capsulatus/metabolism , Animals , Cattle , Dicyclohexylcarbodiimide/metabolism , In Vitro Techniques , Kinetics , Mitochondria, Heart/enzymology , Permeability , Photosynthesis , Protein Conformation , Proton-Translocating ATPases/chemistry , ProtonsABSTRACT
Summary Diazoles have attracted considerable attention for a long time owing to their potentially interesting chemical, biochemical, and medicinal properties. We have reported the synthesis and in vitro antibacterial activity of a new series of (N-substituted)-3-methyl-4-diazo-5-pyrazolecarboxamides 1a-n along with their quantitative structureactivity study. There was a trend towards a better Gram-negative activity with decreasing molecular refraction values of the substituents on the carboxamidic moiety and a better Gram-positive activity with increasing values of IR carbonyl shift. The compounds which displayed the broadest antibacterial spectrum were 3-methyl-4-diazo-5-pyrazolecarboxamides substituted with 2-pyridinyl and 3-isoxazolyl moieties, making evident the structural requirement of a heterocyclic substituent with aminic function located in the ortho position with respect to heteroatom(s). The introduction of new substituents with different lipophilic properties in place of the C(3)-methyl group such as cyano, methoxy, ethoxy, benzyloxy substituents have been examined followed by application of these materials to the cosmetic formulations. An emulsion utilizing the 2-pyridinyl compound at 0.2% and 0.4% has been prepared, and their specific antimicrobial activity has been evaluated. Preliminary antimicrobial challenge studies indicated the excellent preservative activity of this compound in cosmetic formulations.
ABSTRACT
To characterize the role of supernumerary subunits of the mammalian F0F1 ATP synthase, cross-reconstitution of mitochondrial and bacterial F0F1 complexes has been carried out. Escherichia coli F1 (EcF1) can be reconstituted with F1-stripped everted membranes of E. coli (UPEc) and of bovine heart mitochondria (USMP). Bovine heart mitochondrial F1 (BHF1) can also be reconstituted with both membranes. Both EcF1 and BHF1, when reconstituted with UPEc, exhibited oligomycin-insensitive ATP-hydrolase activity. Subunits of the mammalian F0, in particular F0I-PVP protein, F6 and oligomycin-sensitivity-conferring protein (OSCP) conferred oligomycin sensitivity to the catalytic activity of EcF1 or BHF1 reconstituted with UPEc. Reaction of N,N'-dicyclohexylcarbodiimide and development of inhibition of passive H+ conduction was, in UPEc, considerably slower and exhibited a lower apparent affinity than in USMP. The ATP hydrolase activity of UPEc+EcF1 or UPEc+BHF1 was, also, less sensitive to inhibition by N,N'-dicyclohexylcarbodiimide than USMP+EcF1 or USMP+BHF1. Addition of mitochondrial F0I-PVP to UPEc enhanced the sensitivity of H+ conduction to oligomycin. F0I-PVP and OSCP added to UPEc, promoted inhibition by N,N'-dicyclohexylcarbodiimide of passive H+ conduction and increased its binding affinity to subunit c of E. coli F0. The presence of F0I-PVP and OSCP also promoted inhibition by N,N'-dicyclohexylcarbodiimide of the ATP-hydrolase activity of EcF1 or BHF1 reconstituted with UPEc.
Subject(s)
Escherichia coli/enzymology , Mitochondria, Heart/enzymology , Proton-Translocating ATPases/metabolism , Adenosine Triphosphate/metabolism , Animals , Binding Sites , Cattle , Dicyclohexylcarbodiimide/pharmacology , Electrophoresis, Polyacrylamide Gel , Membrane Proteins/metabolism , Oligomycins/metabolism , Oligomycins/pharmacology , ProtonsABSTRACT
Reproductive performance was evaluated after conservative surgery for uterine adenomyoma in a prospective, observational study. Twenty-eight women with histologically proven adenomyomas had conservative surgery in the period 1985-1990. Eighteen patients wanting children were regularly followed for a mean +/- SD period of 53.2 +/- 23.5 months. Crude and cumulative pregnancy rates were calculated after the operation. Thirteen (72.2%) women conceived, for a total of 18 pregnancies: nine (50%) ended in term deliveries, seven (38.8%) in spontaneous abortions, one (5.6%) in ectopic pregnancy and one (5.6%) in a pre-term delivery with neonatal death. The cumulative pregnancy rate at 36 months of follow-up was 74.7%. This analysis of a small series indicates that conservative surgery for adenomyomas is associated with a good reproductive prognosis.