The effect of tactile-kinesthetic stimulation on growth indices of healthy neonates

Abstract Therapeutic touch is emphasized by healthcare professionals for improvement of neonates' growth and development. However, inconsistencies exist regarding effects and methods of massage in neonates. The purpose of this clinical trial is to assess and comprise intervention and control groups regarding the effects of tactile-kinesthetic stimulation (TKS) by mothers on growth indices of healthy term neonates. Sixty healthy term neonates were randomly assigned into intervention and control groups. Mothers of neonates in the experimental group were trained to perform TKS for their newborns at home before feeding for at least 28 consecutive days, two times a day, and 15 min each time. Neonates in the control group were not required to receive this intervention. The neonates’ growth indices were measured within 24 h after birth, and then at days 14 and 28. During the study and the three consecutive measurements, no significant difference was found between the mean weights, heights, and head circumferences of the neonates in the two groups (p > 0.05). Keywords: Tactile-kinesthetic stimulation Growth indices Neonates Mother's touc

The response of the different soybean metallothionein isoforms to cadmium intoxication

Cadmium is a highly toxic heavy metal for both plants and animals. The presence of Cd in agricultural soils is of major concern regarding its entry into the food chain, since Cd compounds are readily taken up by plants, and accumulated in edible parts due to their high solubility. In this study, we first demonstrate the high capacity for Cd concentration of soybean grains. Consequently, we considered the study and characterization of the molecular determinants of Cd accumulation -such as metallothioneins (MT)- to be of major practical importance. We report here the first characterization of the soybean MT system, with the identification of nine genes (one of which is a truncated pseudogene), belonging to the four plant MT types. The most highly expressed of each type was chosen for further function analysis. All of them are expressed at high levels in soybean tissues: GmMT1, GmMT2 and GmMT3 in roots, shoots and seeds, and GmMT4 only in seeds. The corresponding recombinant soybean MTs, synthesized in Escherichia coli cells cultured in metal supplemented media, exhibit greater cadmium than zinc binding capacity. These results suggest a definite role of GmMTs in Cd(II) accumulation as one of the main responses of soybean to an overload of this metal.Fil: Pagani, María Ayelén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Tomas, M.. Universidad de Barcelona; España. Universitat Autònoma de Barcelona; EspañaFil: Carrillo, Julieta Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Bofill, R.. Universitat Autònoma de Barcelona; EspañaFil: Capdevila, M.. Universitat Autònoma de Barcelona; EspañaFil: Atrian, S. Universidad de Barcelona; EspañaFil: Andreo, Carlos Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentin

Cloning, expression and characterization of alcohol dehydrogenases in the silkworm Bombyx mori

Alcohol dehydrogenases (ADH) are a class of enzymes that catalyze the reversible oxidation of alcohols to corresponding aldehydes or ketones, by using either nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP), as coenzymes. In this study, a short-chain ADH gene was identified in Bombyx mori by 5′-RACE PCR. This is the first time the coding region of BmADH has been cloned, expressed, purified and then characterized. The cDNA fragment encoding the BmADH protein was amplified from a pool of silkworm cDNAs by PCR, and then cloned into E. coli expression vector pET-30a(+). The recombinant His-tagged BmADH protein was expressed in E. coli BL21 (DE3), and then purified by metal chelating affinity chromatography. The soluble recombinant BmADH, produced at low-growth temperature, was instrumental in catalyzing the ethanol-dependent reduction of NAD+, thereby indicating ethanol as one of the substrates of BmADH

Characterization of Loading Responses and Failure Loci of a Boron Steel Spot Weld

Boron steel, classed as an ultra high-strength steel (UHSS), has been utilized in anti-intrusion systems in automobiles, providing high strength and weight-saving potential through gage reduction. UHSS spot welds exhibit unique hardness distributions, with a hard nugget and outlying base material, but with a soft heat-affected zone in-between these regions. This soft zone reduces the strength of the weld and makes it susceptible to failure. Due to the interaction of various weld zones that occurs during loading, there is a need to characterize the loading response of the weld for accurate failure predictions. The loading response of certain weld zones, as well as failure loci, was obtained through physical simulation of the welding process. The results showed a significant difference in mechanical behavior through the weld length. An important result is that instrumented indentation was shown to be a valid, quantitative method for verifying the accuracy with which weld microstructure has been recreated with regard to the target weld microstructure

Asimilación metabólica de los alcoholes en drosophzla: nutrición y detoxificación

Alcohol rnetabolism in Drosophila has recently attracted a lot of interest probably due to the essentiai role that the enzyme alcohol dehydrogenase plays in alcohol transforrnation. Species differ markedly in alcohol tolerance and this is attributed to ADH. While some species can cope with high ethanol concentrations (28%), others do not survive when reared in rninimal amounts (0'9%). Most of the species belong to an interrnediate group being able to tolerate and rnake use of alcohols produced in ferrnenting fruits which frequently constitute their natural habitats, al least during embryonic and larval developrnent. The analysis of ethanol metabolisrn and other primary alcohols, will help to establish not only the biochernical transformations, the enzyrnes involved and the relationships between other rnetabolic pathways but also to clarify the various factors relevant to the species adaptation towards alcohol-rich or rnoderately rich environrnents. Different types of evidence seern to indicate that one of these key factors is clearly ADH. In its absence, species become extremely sensitive to ethanol. Yet, it is still unclear which enzyrne features account for higher ethanol tolerance. Avalaible data suggest that the kinetic coeficient Vrnax and the quantity of enzyrne synthesized could be the rnost relevant factors in alcohol adaptation. On the other hand, kinetic and structurai studies of ADH reveal that ethanol utilization probably represents a recent evolutionary acquisition and was not associated initially with the enzyrne. Other enzyrnatic systerns help Drosophila in the exploitation of alcohol-rich environments, although they only account for sorne 10% of alcohol utilization. Aldehydes produced are further oxidized to carboxilic acids and this seems to be produced by an aldehyde dehydrogenase (ALDH), recently characterized in Drosophila, which shares relevant biochemical properties with liver ALDH. Ethanol is then eventually transforrned to acetic acid and acetylCoA which may serve as substrate for fatty acid synthesis or be oxidized in the Krebs cycle. All appears to be different when ADH acts on secondary alcohols. Highly toxic, rnetabolically inert compounds are produced which at the sarne time inactivate the enzyrne. Nevertheless, ADH shows lowest Km values with these substrates. sornething that cannot be easily explained, at least in biological terrns. It has been clairned that greater affínity could be due to the type of chernical interaction between the substrate and the active site of the enzyrne. Studies have also been carried out to know the effect of ethanol and 1 or sucrose in alcohol rnetabolisrn to approach the real situation that Drosophila encounters in natural habitats. It has been proved that ethanol and sucrose, together or individually, exert a positive regulation on ADH activity. It is not clear what produces this effect: increased levels of synthesis of the enzyme, lower turnover rate, re-activation of preexisting less active molecular forrns or if it is directly produced by these compounds or its metabolic interrnediates. The positive regulatory effect of ethanol and sucrose combined only appears if sucrose concentration does not reach a threshold level, when this is exceeded, a negative effect would be produced. When sucrose, at permisible amounts under the inhibitory effect, is supplernented to an ethanol diet, positive rnetabolic regulation is also observed in the activities of lipogenic enzyrnes together with an increase in the triacylglycerol content. Other type of data seem to strenghten the view that ADH role in alcohol transfonnation is not necessarily related to its possible function in intermediary metabolism. Among these are the facts that in the absence of ethanol ADH-negative mutarits differ from wild-type individuals in these metabolic pathways and that sucrose done can modulate ADH activity. In Drosophila , a clear picture of the relationships between catabolic and anabolic pathways has not yet emerged and this also applies to individual pathways. Alcohol metabolism is a clear example: it is not known how the activities of enzymes involved in this process are regulated and if this is acchieved through a direct or indirect effect. Until this stage is reached much of the meaning of the experimental results will be difficult to grasp. Obviously, there is a long way to go before there will be an amount of data comparabre toother organisms, as is the case of mammals, frequently investigated because they serve as model systems for humans.El metabolismo de los alcoholes en Drosophila es objeto de múltiple interés científico, originado por el protagonismo que en él ejerce el enzima alcoholdeshidrogenasa (ADH). En los últimos años, el aporte de nuevos resultados ha sido considerablemente rico y relevante, llegando a poner en duda algunos de los dogmas que durante largo tiempo rigieron este esquema metabólico y proporcionar, por fin, datos sobre otros enzimas implicados. El presente trabajo pretende revisar de modo exhaustivo y critico los datos acumulados hasta la fecha, así como proporcionar una visión integradora de todos ellos en unas vías metabólicas basales e interrelacionadas

The role of elements in the treatment of eye disorders

Background: Three main eye disorders are glaucoma, cataract and age-related macular degeneration. Glaucoma is a progressive, irreversible disease and the second cause of blindness worldwide. This disease involves abnormally high pressure of the fluid inside the eye, which, if left untreated, can result in damage to the optic nerve and vision loss. Cataract is a clouding that develops in the crystalline lens of the eye or in its envelope (lens capsule), results in slight to complete opacity and obstructing the passage of light. Age-related macular degeneration is associated with formation of new blood vessels and leads to vision impairment. Results: Because of the irreversible outcome of eye disease, early treatment of these diseases is very important. Therefore, many attempts are done to find the effective and early treatments (current treatments include eye drops, tablets, laser and other surgery). Surgical treatment is not a cost- and time- effective and associated with many negative side effects. Conclusion: Using elements and materials such as Cerium, inhibitors of Magnesium and Calcium and construction of nano-polymers with organic materials, such as polylactic acid, polylactic/glycolic acid, Hyaluronic acid and acrylic acid can help in treatment of abovementioned disorders. In the present study, at first glaucoma, its causes, role of genetics and then the treatment methods and mechanisms of drugs impact for the disease were reviewed and some drugs were compared with each other

Raman spectroscopy investigation on metal-metallothionein complexes.

Metallothioneins (MTs) are low molecular weight, cysteine-rich proteins with an exceptional heavy metal coordination capacity. Because of their ability to bind metals and to scavenge oxidant radicals, MTs are considered to play a role in metal homeostasis, metal detoxification and control of the oxidative stress. Their high heterogeneity, metal binding abilities and primary structure suggest very diverse functional specializations. Structural and functional studies have been mainly devoted to vertebrate MTs. Participation of metal ligands other than Cys and the presence of secondary structure elements in metal-MT complexes are fairly unknown, especially in non-vertebrate MTs. Recently, it has been shown that other ligands can participate in metal coordination; two main ligand types have been identified: endogenous, as His residues and exogenous, as inorganic ions (i.e. S2- or Cl-). In vivo-synthesized Zn-MTs, representative of different families (mollusc, insect, nematode, echinoderm, vertebrate and plant, enclosing the mammalian MT1 isoform), were heterologously synthesized in E.coli and studied by analytic and spectroscopic techniques. The examined MTs (CeMT2, MeMT, SpMTA, MtnB, QsMT and MT1) contain 43-73 a.a., among which at least a 30% are Cys. To evaluate the influence of the metal on the MT structure, also the Cd-QsMT was synthesised and analysed in comparison with Zn-QsMT. Almost all the MTs considered are devoid of aromatic residues (only Phe in SpMTA and QsMT, Tyr in CeMT2, and His in CeMT2 and QsMT) and contain variable amounts of metal and S2- ions, quantitatively evaluated by acid ICP-AES and GC-FPD measurements. The formation of more than one species (S2--containing and S2--devoid complexes), revealed by ESI-MS spectra, was evident for all the MTs. The Raman spectra gave information about the structure of the metal-MT aggregates. In particular, the presence of secondary structure elements was determined: for the examined MTs isoforms, a relevant contribution of p-sheet and p-turns was shown, whereas the a-helix content resulted almost negligible. As regards Cys sulfurs, almost all Cys were involved in the metal coordination, as indicated by several bands attributable to metal-S stretching modes at low wavenumbers (<500 cm-1). In particular, the high number of vM-S bands, as well as their broadening, suggest the formation of different metal centres. Raman bands markers of sulfide bridging ligands were also identified. In Zn- CeMT2 and Zn-QsMT the eventual participation of the His residue in metal binding was evaluated through a curve fitting analysis of thè 1630-1565 cm"' region. By considering the integrated intensity of the bands, it can be concluded that His residues are mainly coordinated in Zn-CeMT2 whereas in Zn-QsMT His is mainly present as free tautomer. The combination of analytical and spectroscopic techniques has been highly informative for the analysis of in vivo-synthesized metal-MT complexes; Raman studies revealed one of the most promising experimental strategies to provide new data on the knowledge of the metal binding behavior of MTs from the most diverse organisms

Raman study of in vivo synthesized Zn(II)-metallothionein complexes: structural insight into metal clusters and protein folding.

Metallothioneins (MTs) are metal-chelating peptides that play an active role in zinc homeostasis. The participation of metal ligands other than cysteines and the presence of secondary structure elements in metal-MT complexes are fairly unknown, especially in nonvertebrate MTs. Here, four Zn(II) complexes of invertebrate MTs (mollusc, insect, nematode, and echinoderm) and the Zn(II)-MT complex of the mammalian MT1 isoform, heterologously synthesized in E. coli, were studied by analytic and spectroscopic techniques. By Raman and circular dichroism spectroscopy, new structural informations were obtained. The five analyzed MT isoforms consist largely of -turns with the near exclusion of -helical segments. Raman spectroscopy was revealed as an useful tool, providing information about the state of the cysteine sulfur atoms (metal coordinated and oxidized), the participation of histidine in metal coordination, and the molecular environment of tyrosine residues. In all the five Zn(II)-MT studied samples, acid-labile sulfide anions were found as nonproteic ligands, since sulfide-containing and sulfide-devoid species coexisted in the corresponding preparations. Significantly, Raman bands useful as markers of sulfide bridging ligands were identified. Overall, this work illustrates how the combination of analytical and spectroscopic techniques can be a very informative approach for the analysis of in vivo-synthesized metal-MT complexes, providing new data on the metal binding behavior of MTs from the most diverse organisms. \ua9 2008 Wiley Periodicals, Inc. Biopolymers 89: 1114-1124, 2008. This article was originally published online as an accepted preprint. The Published Online date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at [email protected]

Comparative insight into the Zn(II)-, Cd(II)- and Cu(I)-binding features of the protozoan Tetrahymena pyriformis MT1 metallothionein.

Tetrahymena pyriformis MT1 (TpyMT1) is a model among ciliate metallothioneins (MTs). Here, we report on the analytic (ICP-AES, GC-FPD), spectroscopic (CD, UV\u2013Vis, Raman) and spectrometric (ESI-MS) characterization of its recombinant Cd(II)-, Zn(II)- and Cu(I)-complexes, and of those formed during in vitro Zn/Cd and Zn/Cu replacement. In the presence of Cd(II), TpyMT1 renders a major Cd11\u2013TpyMT1 species, which is also the final step reached in the in vitro Zn/Cd exchange process in Zn11\u2013TpyMT1. Spectroscopic data supports a different folding of the isostoichiometric Cd11\u2013 and Zn11\u2013TpyMT1 complexes. Unexpectedly, TpyMT1 biosynthesis in Zn(II)-rich cultures was sensitive to the aeration degree, so that high oxygenation rendered undermetalated, partially-oxidized, complexes (Zn9\u2013TpyMT1). Biosynthesis in Cu(I)-rich media rendered extremely heterogeneous mixtures of CuxZny-species (x + y = 8\u201320), where the higher the aeration, the higher the Zn(II) content. The complexity of these samples was reproduced during the Zn/Cu replacement, as the number of generated species increased gradually with the addition of copper to Zn11\u2013TpyMT1. According to our results, a clear preference of TpyMT1 for Cd(II) binding, rather than for Zn(II), and especially Cu(I) can be postulated. This character is totally consistent with the induction pattern of the TpyMT1 gene and the postulated role of TpyMT1 in Cd-detoxification