The primary structure of the flavoprotein D-aspartate oxidase from beef kidney.

The complete primary structure of the peroxisomal flavoenzyme D-aspartate oxidase from beef kidney has been determined by analyses of the peptides obtained through fragmentation of the carboxymethylated protein with trypsin, CNBr, heptafluorobutyric acid/CNBr and Staphylococcus aureus V8 protease. The protein consists of a single polypeptide of 338 residues, accounting for a M(r) of 37,305 for the apoprotein. A form of the enzyme lacking Lys-338 and therefore ending with Pro-337 has been detected. The N-terminal residue is blocked. Seven cysteines and no disulfide bridges are present. Residue 228 can be either Ile or Val. Thus, D-aspartate oxidase presents two types of heterogeneity in the polypeptide chain in addition to the one already described concerning the possible content of FAD or 6-hydroxyflavin adenine dinucleotide. Comparison of the primary structure of D-aspartate oxidase with other known sequences reveals that D-aspartate oxidase is homologous with D-amino acid oxidase (another flavo-oxidase) and does not present significant sequence similarities with any other protein, including flavoenzymes

Cosmetics for acne: indications and recommendations for an evidence-based approach.

AIM: The aim of this review was to evaluate, by a thorough revision of the literature, the true efficacy of currently available topic and systemic cosmetic acne agents. METHODS: The efficacy of currently available cosmetic acne agents has been retrospectively evaluated via thorough revision of the literature on matched electronic databases (PubMed). All retrieved studies, either randomized clinical trials or clinical trials, controlled or uncontrolled were considered. RESULTS: Scientific evidence suggests that most cosmetic products for acne may enhance the clinical outcome. Cleansers should be indicated to all acne patients; those containing benzoyl peroxide or azelaic/salicylic acid/triclosan show the best efficacy profile. Sebum-controlling agents containing nicotinamide or zinc acetate may minimize excessive sebum production. Cosmetics with antimicrobial and anti-inflammatory substances such as, respectively, ethyl lactate or phytosphingosine and nicotinamide or resveratrol, may speed acne recovery. Topical corneolytics, including retinaldehyde/glycolic acid or lactic acid, induce a comedolytic effect and may also facilitate skin absorption of topical drugs. Finally, the use of specific moisturizers should be strongly recommended in all acne patients. CONCLUSION: Cosmetics, if correctly prescribed, may improve the performance of the therapy, whereas wrong procedures and/or inadequate cosmetics may worsen acne. Cosmetological recommendations may allow clinicians to make informed decisions about the role of various cosmetics and to indentify the appropriate indications and precautions. The choice of the most effective product should take into consideration the ongoing pharmacological therapy and acne type/severity as well

Kombinirano liječenje makularnog edema uzrokovanog okluzijom mrežnične vene bevacizumabom i triamcinolon acetonidom

The purpose of this study was to determine the efficacy of combined intravitreal bevacizumab and triamcinolone in the treatment of macular edema due to retinal vein occlusion. A prospective randomized trial was conducted in the Department of Ophthalmology, Osijek University Hospital Centre in Osijek including 51 patients divided into three groups depending on the drug received. The first group received 1.25 mg intravitreal bevacizumab, the second group received 1 mg intravitreal triamcinolone, and the third group received a combination of 1.25 mg bevacizumab and 1 mg intravitreal triamcinolone on the same day. Changes in the central macular thickness, intraocular pressure and visual acuity were monitored during the follow up period. The retinal perfusion status was evaluated by fluorescein angiography. The group that received combined treatment had better outcome in terms of reduction of macular thickness. There was no statistically significant intraocular pressure elevation among the three treatment groups or within each group of patients. A positive trend regarding visual improvement was observed in the group receiving combined treatment in spite of the lowest initial visual acuity, highest value of macular thickness and longest mean duration of symptoms. In conclusion, combined treatment with bevacizumab and triamcinolone for the treatment of retinal vein occlusion is more potent, safe, efficient and cost-effective. It can also be recommended because fewer injections are needed in patients undergoing treatment for macular edema.Cilj rada bio je utvrditi učinkovitost kombinirane intravitrealne terapije bevacizumabom i triamcinolonom kod makularnog edema nastalog kao posljedica okluzije mrežnične vene. Prospektivno randomizirano ispitivanje provedeno je na Odjelu za očne bolesti Kliničkoga bolničkog centra Osijek. U ispitivanje je bio uključen 51 ispitanik, koji su podijeljeni u tri skupine ovisno o vrsti lijeka koji su primali. Bolesnici u prvoj skupini primali su 1,25 mg bevacizumaba intravitrealno, druga skupina ispitanika je primala 1 mg triamcinolona intravitrealno, a treća skupina je primala kombinaciju 1,25 mg bevacizumaba i 1 mg triamcinolona intravitrealno u istom posjetu. Tijekom razdoblja praćenja promatrane su promjene u centralnoj makularnoj debljini, vidnoj oštrini, kao i vrijednosti intraokularnog tlaka. Fluoresceinska angiografija primijenjena je za procjenu perfuzijskog statusa retine. Skupina koja je primila kombiniranu terapiju s oba lijeka imala je bolji ishod u vidu smanjenja makularne debljine. Nije bilo značajnijeg povišenja očnog tlaka unutar skupina, kao ni usporedbom među skupinama. U skupini ispitanika koji su primili kombinaciju oba lijeka zabilježen je pozitivan trend u oporavku vidne oštrine, iako su imali najniže ulazne vrijednosti vidne oštrine, najveću vrijednost centralne makularne debljine mjerenu optičkom koherentnom tomografijom i najduže prosječno trajanje okluzije u odnosu na ostale skupine ispitanika. Zaključno, smanjenje broja injekcija kod primjene kombinacije oba lijeka predstavlja ekonomičniji pristup liječenju okluzije mrežnične vene, a također djeluje potentnije na sniženje centralne makularne debljine u odnosu na pojedinačnu primjenu svakog lijeka

A novel promising laccase from the psychrotolerant and halotolerant Antarctic marine Halomonas sp. M68 strain

Microbial communities inhabiting the Antarctic Ocean show psychrophilic and halophilic adaptations conferring interesting properties to the enzymes they produce, which could be exploited in biotechnology and bioremediation processes. Use of cold- and salt-tolerant enzymes allows to limit costs, reduce contaminations, and minimize pretreatment steps. Here, we report on the screening of 186 morphologically diverse microorganisms isolated from marine biofilms and water samples collected in Terra Nova Bay (Ross Sea, Antarctica) for the identification of new laccase activities. After primary screening, 13.4 and 10.8% of the isolates were identified for the ability to oxidize 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and the dye azure B, respectively. Amongst them, the marine Halomonas sp. strain M68 showed the highest activity. Production of its laccase-like activity increased six-fold when copper was added to culture medium. Enzymatic activity-guided separation coupled with mass spectrometry identified this intracellular laccase-like protein (named Ant laccase) as belonging to the copper resistance system multicopper oxidase family. Ant laccase oxidized ABTS and 2,6-dimethoxy phenol, working better at acidic pHs The enzyme showed a good thermostability, with optimal temperature in the 40–50°C range and maintaining more than 40% of its maximal activity even at 10°C. Furthermore, Ant laccase was salt- and organic solvent-tolerant, paving the way for its use in harsh conditions. To our knowledge, this is the first report concerning the characterization of a thermo- and halo-tolerant laccase isolated from a marine Antarctic bacterium

Low n-6/n-3 Gestation and Lactation Diets Influence Early Performance, Muscle and Adipose Polyunsaturated Fatty Acid Content and Deposition, and Relative Abundance of Proteins in Suckling Piglets

Elevated omega-6 (n-6) and omega-3 (n-3) polyunsaturated fatty acids (PUFAs) ratios in swine diets can potentially impose a higher risk of inflammatory and metabolic diseases in swine. A low ratio between the two omega PUFAs has beneficial effects on sows' and piglets' production performance and immunity status. At present, there are few studies on how sow nutrition directly affects the protein and fat deposition in suckling piglets. Two groups of sows were fed diets with high or low n-6/n-3 polyunsaturated ratios of 13:1 (SOY) and 4:1 (LIN), respectively, during gestation and lactation. Longissimus dorsi muscle and adipose tissue from newborn piglets, nourished only with sow's milk, were subjected to fatty acid profiling by gas chromatography-mass spectrometry (GC-MS) and to proteomics assays based on nano-liquid chromatography coupled to high-resolution tandem mass spectrometry (nLC-HRMS). Fatty acid profiles on both muscle and adipose tissues resembled the magnitude of the differences between fatty acid across diets. Proteomic analysis revealed overabundance of 4 muscle and 11 adipose tissue proteins in SOY compared to LIN in both piglet tissues. The detected overabundance of haptoglobin, an acute-phase protein, and the stimulation of protein-coding genes and proteins related to the innate immune response and acute inflammatory response could be associated with the pro-inflammatory role of n-6 PUFAs

Cognitive dysfunctions and psychological symptoms in migraine without aura: a cross-sectional study

BACKGROUND: The occurrence of cognitive dysfunctions and psychological symptoms, as well as their mutual relationships, in migraine patients are still debated. The aim of the study was to characterize the cognitive profile and psychological symptoms (i.e. depression, anxiety and apathy) in drug-naïve migraine without aura (MwoA) patients. METHODS: Seventy-two consecutive MwoA patients, referred to the Italian University Headache Clinic and 72 healthy subjects (HCs) were enrolled. Patients, during an attack-free period, and HCs completed Montreal Cognitive Assessment (MoCA), Beck Depression Inventory-II (BDI-II), Self-version of Apathy Evaluation Scale (AES-S) and State and Trait Anxiety Inventory (STAI-Y-1 and 2). Clinical parameters of disease severity (i.e. disease duration, migraine attacks per month, mean pain intensity during migraine attacks, migraine disability and impact on daily life) were recorded. RESULTS: Although performance of MwoA patients on MoCA was above Italian cut-off threshold (<15.5) suggesting presence of cognitive impairment, MwoA patients achieved significantly lower scores than HCs on total MoCA scale (22.3 ± 2.7 versus 25.4 ± 2.3) and on its attention (4.9 ± 1.1 versus 5.6 ± 0.7), memory (1.8 ± 1.4 versus 3.1 ± 1.3), visuospatial (3.2 ± 0.9 versus 3.6 ± 0.6) and executive subscales (2.6 ± 1.1 versus 3.1 ± 0.8). In addition, we observed significant correlations between MoCA executive domain subscore and the attack-related disability score (MIDAS). As for behavioral profile, the percentage of depressive symptoms (4.2 %), high state and trait anxiety (13.9 and 9.7 %, respectively), and apathy (11.1 %) in MwoA patients were similar to that of HCs. No significant associations of behavioural symptoms with cognitive performance and clinical parameters were found. CONCLUSIONS: Drug-naïve MwoA patients are characterized by subtle cognitive dysfunctions and low percentage of behavioural symptoms. The results support the importance of searching for subclinical cognitive disturbances in patients with MwoA, who deserve to be followed-up to verify whether they develop clinically relevant disorders over time

Limited proteolysis of a disulfide-linked apoA-I dimer in reconstituted HDL.

The apolipoprotein A-I Milano (apoA-I M ) is a mo- lecular variant of apoA-I characterized by the Arg 173 → Cys substitution, leading to the formation of homodimers A-I M / A-I M. Upon interaction with palmitoyloleoylphosphatidyl- choline, A-I M /A-I M forms only two species of reconstituted HDL (rHDL) particles, with diameters of 7.8 and 12.5 nm. We used limited proteolysis to analyze the conformation of A-I M /A-I M in the two rHDL particles, in comparison with that of apoA-I in rHDL of similar size. ApoA-I in the small, 7.8-nm rHDL is degraded to a greater extent (50% after 6 h) than in the large rHDL ( � 10% degraded after 6 h). The pro- tease susceptibility of A-I M /A-I M in small and large rHDL is instead remarkably the same, with A-I M /A-I M being much more sensitive to proteolytic digestion (50% degraded after 10 min) than apoA-I. The identification of the proteolytic fragments by immunoblotting, N-terminal sequencing, and molecular mass determination, shows that the N-terminus of both proteins is resistant to proteolysis, with six cleavage sites located in the central and carboxy-terminal portions of the molecules. Cleavage in the middle of apoA-I occurs at dis- tinct sites in 7.8-nm (Lys 118 ) and 12.7-nm (Arg 123 ) rHDL, in- dicating a different conformation in small and large rHDL particles. The A-I M /A-I M instead adopts a unique and identi- cal conformation in small and large rHDL, with the carboxy- terminal portion of the molecule being remarkably more ac- cessible to the proteases than in apoA-I. This suggests the presence of a novel carboxy-terminal domain in A-I M /A-I M , not organized in a compact structure and not shared by wild-type apoA-I, which may account for the unique functional proper- ties of A-I M /A-I M. —Calabresi, L., G. Tedeschi, C. Treu, S. Ron- chi, D. Galbiati, S. Airoldi, C. R. Sirtori, Y. Marcel, and G. Franceschini. Limited proteolysis of a disulfide-linked apoA-I dimer in reconstituted HDL. J. Lipid Res. 2001. 42: 935-942

Conversion of nanoscale topographical information of cluster-assembled zirconia surfaces into mechanotransductive events promotes neuronal differentiation

Additional file 4: Table S1. Proteomic data for upregulated proteins. Proteins upregulated (compared to flat-Zr) or present only in cells grown on ns-Zr15. Adhesome proteins and proteins with roles in mechanobiological processes are marked in dark and light grey, respectively

Serine metabolism during differentiation of human iPSC-derived astrocytes

: Astrocytes are essential players in development and functions, being particularly relevant as regulators of brain energy metabolism, ionic homeostasis and synaptic transmission. They are also the major source of l-serine in the brain, which is synthesized from the glycolytic intermediate 3-phosphoglycerate through the phosphorylated pathway. l-Serine is the precursor of the two main co-agonists of the N-methyl-d-aspartate receptor, glycine and d-serine. Strikingly, dysfunctions in both l- and d-serine metabolism are associated with neurological and psychiatric disorders. Here, we exploited a differentiation protocol, based on the generation of human mature astrocytes from neural stem cells, and investigated the modification of the proteomic and metabolomic profile during the differentiation process. We show that differentiated astrocytes are more similar to mature rather than to reactive ones, and that axogenesis and pyrimidine metabolism increase up to 30 days along with the folate cycle and sphingolipid metabolism. Consistent with the proliferation and cellular maturation processes that are taking place, also the intracellular levels of l-serine, glycine, threonine, l- and d-aspartate (which level is unexpectedly higher than that of d-serine) show the same biosynthetic time course. A significant utilization of l-serine from the medium is apparent while glycine is first consumed and then released with a peak at 30 days, parallel to its intracellular level. These results underline how metabolism changes during astrocyte differentiation, highlight that d-serine synthesis is restricted in differentiated astrocytes and provide a valuable model for developing potential novel therapeutic approaches to address brain diseases, especially the ones related to serine metabolism alterations

Proteomic/peptidomic profile and Escherichia coli growth inhibitory effect of in vitro digested soya protein

Plant proteins contain bioactive peptides with functional properties and physiological activities. In the present work, the bioactive peptides produced during in vitro gastrointestinal digestion of soya protein isolate were investigated. Protein samples were subjected to simulated gastrointestinal digestion with a generation of permeate (3 kDa) fractions. The permeate was analysed by nano-liquid chromatography electrospray ionisation tandem mass spectrometry (LC-nano ESI MS/MS) using a shotgun peptidomic approach, and the retentate was further digested with trypsin and analysed using a shotgun proteomic approach. Based on protein profile observed, the retentate was further tested for its potential antimicrobial activity by evaluating the inhibitory effect on E. coli growth. In the present study the peptidomic/ proteomic characterisation of permeate and retentate fractions revealed the presence of bioactive peptides and proteins associated with antioxidant, ACE-inhibitory, anti-hypertensive and antimicrobial activities. The presence of potentially antimicrobial proteins in the retentate fraction is supported by a marked E. coli F18þ growth inhibitory activity of the same fraction. In particular, the growth inhibitory effect was significant from one until six hours of incubation with 0.65–2.6 mg/ml of in vitro digested soya. The obtained data confirmed that soya-based supplements may have potential beneficial effects after human consumption, and they may be recycled for animal nutrition in line with the circular economy concept