Effect of magnesium chloride and guanidinum chloride on the extraction of components of extracellular matrix from chicken cartilage

In order to evaluate the effect of chaotropic agents on proteoglycan and non-collagenous proteins, chicken xiphoid cartilage was treated with guanidine-HCI and MgCl2 in different concentrations (1M to 5M), and different periods of time (12, 24, 48 and 72hr). The maximum yield of uronic acid was obtained with 3M MgCl2 (73.3 per cent). Concentrations of 4M and 5M of MgCl2 showed that much less uronic acid was removed, 55.3 per cent and 38.1 respectively. Extraction with 3M MgCl2 and 3M guanidine-HCl resulted better efficiency when performed for 48 hr. Analysis by SDS-PAGE of the extracts obtained with guanidine-HCl and MgCl, in different concentrations pointed out that most components are equally removed with the two solvents, showing that the extraction with MgCl2 is an alternative assay to remove non-collagenous proteins from extracellular matrix.

Magnesium chloride alone or in combination with diazepam fails to prevent hippocampal damage following transient forebrain ischemia

In the central nervous system, magnesium ion (Mg2+) acts as an endogenous modulator of N-methyl-D-aspartate (NMDA)-coupled calcium channels, and may play a major role in the pathomechanisms of ischemic brain damage. In the present study, we investigated the effects of magnesium chloride (MgCl2, 2.5, 5.0 or 7.5 mmol/kg), either alone or in combination with diazepam (DZ), on ischemia-induced hippocampal cell death. Male Wistar rats (250-300 g) were subjected to transient forebrain ischemia for 15 min using the 4-vessel occlusion model. MgCl2 was applied systemically (sc) in single (1x, 2 h post-ischemia) or multiple doses (4x, 1, 2, 24 and 48 h post-ischemia). DZ was always given twice, at 1 and 2 h post-ischemia. Thus, ischemia-subjected rats were assigned to one of the following treatments: vehicle (0.1 ml/kg, N = 34), DZ (10 mg/kg, N = 24), MgCl2 (2.5 mmol/kg, N = 10), MgCl2 (5.0 mmol/kg, N = 17), MgCl2 (7.5 mmol/kg, N = 9) or MgCl2 (5 mmol/kg) + DZ (10 mg/kg, N = 14). Seven days after ischemia the brains were analyzed histologically. Fifteen minutes of ischemia caused massive pyramidal cell loss in the subiculum (90.3%) and CA1 (88.4%) sectors of the hippocampus (P<0.0001, vehicle vs sham). Compared to the vehicle-treated group, all pharmacological treatments failed to attenuate the ischemia-induced death of both subiculum (lesion: 86.7-93.4%) and CA1 (lesion: 85.5-91.2%) pyramidal cells (P>0.05). Both DZ alone and DZ + MgCl2 reduced rectal temperature significantly (P<0.05). No animal death was observed after drug treatment. These data indicate that exogenous magnesium, when administered systemically post-ischemia even in different multiple dose schedules, alone or with diazepam, is not useful against the histopathological effects of transient global cerebral ischemia in rats.

Antinociceptive potency of aminoglycoside antibiotics and magnesium chloride: a comparative study on models of phasic and incisional pain in rats

A close relationship exists between calcium concentration in the central nervous system and nociceptive processing. Aminoglycoside antibiotics and magnesium interact with N- and P/Q-type voltage-operated calcium channels. In the present study we compare the antinociceptive potency of intrathecal administration of aminoglycoside antibiotics and magnesium chloride in the tail-flick test and on incisional pain in rats, taken as models of phasic and persistent post-surgical pain, respectively. The order of potency in the tail-flick test was gentamicin (ED50 = 3.34 µg; confidence limits 2.65 and 4.2) > streptomycin (5.68 µg; 3.76 and 8.57) = neomycin (9.22 µg; 6.98 and 12.17) > magnesium (19.49 µg; 11.46 and 33.13). The order of potency to reduce incisional pain was gentamicin (ED50 = 2.06 µg; confidence limits 1.46 and 2.9) > streptomycin (47.86 µg; 26.3 and 87.1) = neomycin (83.17 µg; 51.6 and 133.9). The dose-response curves for each test did not deviate significantly from parallelism. We conclude that neomycin and streptomycin are more potent against phasic pain than against persistent pain, whereas gentamicin is equipotent against both types of pain. Magnesium was less potent than the antibiotics and effective in the tail-flick test only.

Optimization of random amplified polymorphic DNA techniques for use in genetic studies of Cuban triatominae

Random amplified polymorphic DNA (RAPD) technique is a simple and reliable method to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of assay. In this study, we analyzed the effect of changing the concentration of primer, magnesium chloride, template DNA and Taq DNA polymerase with the objective of determining their optimum concentration for the standardization of RAPD technique for genetic studies of Cuban Triatominae. Reproducible amplification patterns were obtained using 5 pmoL of primer, 2.5 mM of MgCl2, 25 ng of template DNA and 2 U of Taq DNA polymerase in 25 µL of the reaction. A panel of five random primers was used to evaluate the genetic variability of T. flavida. Three of these (OPA-1, OPA-2 and OPA-4) generated reproducible and distinguishable fingerprinting patterns of Triatominae. Numerical analysis of 52 RAPD amplified bands generated for all five primers was carried out with unweighted pair group method analysis (UPGMA). Jaccard's Similarity Coefficient data were used to construct a dendrogram. Two groups could be distinguished by RAPD data and these groups coincided with geographic origin, i.e. the populations captured in areas from east and west of Guanahacabibes, Pinar del Río. T. flavida present low interpopulation variability that could result in greater susceptibility to pesticides in control programs. The RAPD protocol and the selected primers are useful for molecular characterization of Cuban Triatominae.

Study on thermostabilizers for trivalent oral poliomyelitis vaccine

Different formulations of trivalent oral poliomyelitis vaccine were tested, in order to obtain better thermostability, reduce corrosion of machinery and improve production costs. Magnesium chloride, sucrose, arginine and 199-Hank's medium were used in the formulations. The most appropriate formulation was a mixture of MgCl2 and arginine, which was highly thermostable, and had low production costs. The low corrosive formulation was rejected, due to low thermostability on storage.

Sweet grape mini tomato grown in culture substrates and effluent with nutrient complementation

The objective of this study was to evaluate production of Sweet Grape mini tomato (Lycopersicum esculentum Mill.) using culture substrates and nutrient solution sewage effluent, applied by drip irrigation (fertigation). The experiment was conducted at the University of Goiás State (UEG-UnUCET), from June to November 2011 in Anápolis-GO, Brazil. The experimental design was a 2 x 3 factorial arrangement in a randomized complete block design with four repetitions. The plots were made by combining two nutrient solutions, effluent supplemented with mineral fertilizers (EcS); conventional nutrient solution (SnC); in addition three cultivation substrates: 60% of fine sand washed + 40% substrate composed by 20% coconut fiber plus 80% pine bark (S1); 20% coconut fiber and 80% pine bark (S2) and natural coconut fiber (S3). Sewage effluent were determined nitrate, calcium, potassium, manganese, total phosphate, total iron, magnesium, chloride, sulphate, boron, zinc and molybdenum. We evaluated average mass and average number of fruits per bunch, total fruit and total yield per plant. Statistical difference absence among tested solutions indicates sewage effluent can be used as an alternative source of nutrients in growing mini tomatoes in hydroponics.

Desiccation tolerance of Tapirira obtusa seeds collected from different environments

This study was aimed at evaluating the desiccation sensitivity in seeds of the tree Tapirira obtusa (Benth.) J. D. Mitchell collected from three different environments and subjected to two distinct drying speeds. Seeds were collected from a rocky area, in the "Cerrado", and in a riparian forest area, in the region of municipality of Lavras, State of Minas Gerais. The seeds were subjected to drying with magnesium chloride (slow drying) or silica gel (fast drying), into closed environment, until moisture contents of 40%, 30%, 20% and 10%, considering as control, the percentage of germination at the initial moisture content in each environment, which varied from 47% to 50%. Percentages of germination and normal seedlings as well as germination speed index were assessed. For the three environments studied, there was no effect of slow drying on seed germination. Seeds from area of Cerrado, however, have shown a slight reduction on germination when subjected to fast drying. Oppositely, seeds from rocky area had germination increased when subjected to fast drying. Seeds from riparian forest area had no reduction on germination percentage, independent of drying speed. Results suggest that seeds of T. obtusa are not sensitive to desiccation.

Effectiveness of fused magnesium potassium phosphate for marandu grass

The current high price of KCl and great dependence on importation to satisfy the Brazilian demand indicate the need for studies that evaluate the efficiency of other K sources, particularly those based on domestic raw material. For this purpose, a greenhouse experiment was conducted with samples of a sandy clay loam Typic Haplustox, in a completely randomized 4 x 3 x 2 factorial design: four K rates (0, 60, 120, and 180 mg kg-1), three sources (potassium chloride (KCl), fused magnesium potassium phosphate (FMPP) and a mixture of 70 % FMPP + 30 % KCl) and two particle sizes (100 and 60 mesh), with three replications. Potassium fertilization resulted in significant increases in shoot dry matter production and in K concentrations, both in soil and plants. The K source and particle size had no significant effect on the evaluated characteristics. Potassium critical levels in the soil and the shoots were 1.53 mmol c dm-3 and 19.1 g kg-1, respectively.

Influence of variables on centrifuge-flotation technique for recovery of Toxocara canis eggs from soil

The purpose of this study was to evaluate the influence of variables in a flotation technique for the recovery of Toxocara canis eggs from soil. The trials were done under standardized conditions on one gram of previously sterilized soil samples contaminated with 200 eggs of T. canis. The following variables were evaluated in serial steps: sieving; type of wash; time of stirring; resuspension of sediment; solution flotation. Centrifuge-flotation in sodium nitrate (d = 1.20 g/cm³) was adopted as an initial technique, using Tween 80 (0.2%) and decinormal sodium hydroxide as solutions for washing the samples. Ten tests were done to compare the variables, using counting in triplicate. The sieving of the material reduced significantly the recovery of eggs (p < 0.001) and the number of eggs recovered was higher when the sediment was resuspended (p < 0.05). After standardization, flotation solutions sodium chloride, zinc sulfate, sodium dichromate, magnesium sulfate, and sodium nitrate (d = 1.20g/cm³) were compared. The best results were obtained by using zinc sulfate solution. In conclusion, the chances of recovering T. canis eggs from samples using flotation solutions can be increased by washing of soil twice using distilled water, and resuspension of sediment. On the other hand, the sieving procedure can drastically reduce the number of eggs.

Cystic fibrosis with normal sweat chloride concentration: case report

Cystic fibrosis is a genetic disease usually diagnosed by abnormal sweat testing. We report a case of an 18-year-old female with bronchiectasis, chronic P. aeruginosa infection, and normal sweat chloride concentrations who experienced rapid decrease of lung function and clinical deterioration despite treatment. Given the high suspicion ofcystic fibrosis, broad genotyping testing was performed, showing a compound heterozygous with deltaF508 and 3849+10kb C->T mutations, therefore confirming cystic fibrosis diagnosis. Although the sweat chloride test remains the gold standard for the diagnosis of cystic fibrosis, alternative diagnostic tests such as genotyping and electrophysiologic measurements must be performed if there is suspicion of cystic fibrosis, despite normal or borderline sweat chloride levels.

Bethanecol chloride for treatment of clomipramine-induced orgasmic dysfunction in males

PURPOSE: To investigate whether bethanecol chloride may be an alternative for the clinical management of clomipramine-induced orgasmic dysfunction, reported to occur in up to 96% of male users. METHODS: In this study, 12 fully remitted panic disorder patients, complaining of severe clomipramine-induced ejaculatory delay, were randomly assigned to either bethanecol chloride tablets (20 mg, as needed) or placebo in a randomized, double-blind, placebo-controlled, two-period crossover study. A visual analog scale was used to assess severity of the orgasmic dysfunction. RESULTS: A clear improvement was observed in the active treatment period. No placebo or carry-over effects were observed. CONCLUSION: These findings suggest that bethanecol chloride given 45 minutes before sexual intercourse may be useful for clomipramine-induced orgasmic dysfunction in males.

Sodium chloride added to transport water and physiological responses of Matrinxã Brycon amazonicus (Teleost: Characidae)

The addition of salt to the water has been used to mitigate stress and improve survival in fishes. This study investigated the effects of sodium chloride (0.0, 1.0, 3.0 and 6.0 g/l) on levels of plasma cortisol, glucose, tryacilglycerol, total protein, hematocrit, hemoglobin, erythrocyte number, liver glycogen and lipid, and muscle lipid in adult matrinxã(Brycon amazonicum) after a 4-h transport and during a 96-h recovery period. Fish were sampled before and after transport, and 24 and 96 h of the recovery period. Plasma cortisol was higher than initial condition immediately after transportation, except in fish transported in 3.0 and 6.0 g/l of salt. A similar pattern was observed for blood glucose but fish transported in water with 0.0, 1.0 and 3.0 g/l of salt needed more than 24 h to return to the initial condition. Liver glycogen was lower after transport in fish not exposed to salt. Hemoglobin, erythrocyte number, total plasma protein and liver lipid did not change during the experiment but hematocrit was lower after transport in all treatments and returned to pre-transport values in 24 h. Reductions of muscle lipid and plasma tryacilglycerol were observed during the recovery period in fish from all treatments. The results show that 6.0 g/l NaCl added to the transport water reduce the stress responses and a 96-h recovery period is needed if no salt is used to mitigate the stress.

Chemical and physical attributes and calcium, magnesium, potassium, and phosphorus in the soil of murundu fields in Brazil

Fields of murundus (FM) are wetlands that provide numerous ecosystem services. The objectives of this study were to evaluate the chemical [organic carbon (OC), P, K+, Ca2+, Mg2+, Al3+ and H+Al] and physical [texture and bulk density (Bd)] soil attributes and calculate the organic matter (OM) and nutrient stock (P, Ca, Mg, and K) in soils of FM located in the Guapore River basin in Mato Grosso. Thirty-six sampling points were selected, and soil samples were collected from two environments: the murundu and plain area surrounding (PAS). At each sampling point, mini trenches of 0.5 × 0.5 × 0.4 m were opened and disturbed and undisturbed soil samples were collected at depths of 0-0.1, 0.1-0.2, and 0.2-0.4 m. In the Principal Component Analysis the variables H+Al (49%) and OM (4%) were associated with the F1 component and sand content (47%) with the F2 component. The FM had lower pH values and higher concentrations of K+, P, and H+Al than PAS at all depths (p < 0.05). Additionally, FM stocked up to 433, 360, 205, and 11 kg ha-1 of Ca, Mg, K, and P, respectively, for up to a depth of 0.2 m. The murundu stored two times more K and three times more P than that in the PAS. Our results show that the FM has high sand content and Bd greater than 1.5 Mg m-3, high acidity, low OC content, and low nutrient concentrations. Thus, special care must be taken to preserve FM such that human intervention does not trigger environmental imbalances.

Late Outcome of a Randomized Study on Oral Magnesium for Premature Complexes

Background:Ventricular and supraventricular premature complexes (PC) are frequent and usually symptomatic. According to a previous study, magnesium pidolate (MgP) administration to symptomatic patients can improve the PC density and symptoms.Objective:To assess the late follow-up of that clinical intervention in patients treated with MgP or placebo.Methods:In the first phase of the study, 90 symptomatic and consecutive patients with PC were randomized (double-blind) to receive either MgP or placebo for 30 days. Monthly follow-up visits were conducted for 15 months to assess symptoms and control electrolytes. 24-hour Holter was performed twice, regardless of symptoms, or whenever symptoms were present. In the second phase of the study, relapsing patients, who had received MgP or placebo (crossing-over) in the first phase, were treated with MgP according to the same protocol.Results:Of the 45 patients initially treated with MgP, 17 (37.8%) relapsed during the 15-month follow-up, and the relapse time varied. Relapsing patients treated again had a statistically significant reduction in the PC density of 138.25/hour (p < 0.001). The crossing-over patients reduced it by 247/hour (p < 0.001). Patients who did not relapse, had a low PC frequency (3 PC/hour). Retreated patients had a 76.5% improvement in symptom, and crossing-over patients, 71.4%.Conclusion:Some patients on MgP had relapse of symptoms and PC, indicating that MgP is neither a definitive nor a curative treatment for late follow-up. However, improvement in the PC frequency and symptoms was observed in the second phase of treatment, similar to the response in the first phase of treatment.

A absorção de nitrogênio, fósforo, potássio, cálcio, magnésio, enxôfre e silício pela cana de açúcar, Co 419, e o seu crescimento em função da idade

This paper describes the data obtained for the growth of sugar cane, Variety Co 419, and the amount and rate of absorption of nitrogen, phosphorus, potassium, calcium, magnesium, sulfur, and silicon, according to the age of the plant, in the soil and climate conditions of the state of S. Paulo, Brazil. An experiment was installed in the Estação Experimental de Cana de Açúcar "Dr. José Vizioli", at Piracicaba, state of S. Paulo, Brazil, and the soil "tèrra-roxa misturada" presented the following composition: Sand (more than 0,2 mm)........................................................................ 8.40 % Fine sand (from 0,2 to less than 0,02 mm)................................................. 24.90 % Silt (from 0,02 to less than 0,002 mm)...................................................... 16.40 % Clay (form 0,002 mm and less)................................................................ 50.20 % pH 10 g of soil and 25 ml of distilled water)..................................................... 5.20 %C (g of carbon per 100 g of soil)................................................................. 1.00 %N (g of nitrogen per 100 g of soil)............................................................... 0.15 P0(4)-³ (me. per 100 g of soil, soluble in 0,05 normal H2SO4) ............................... 0.06 K+ (exchangeable, me. per 100 g of soil)....... 0.18 Ca+² (exchangeable, me. per 100 g of soil)...... 2.00 Mg+² (exchangeable, me. per 100 g of soil)...... 0.66 The monthly rainfall and mean temperature from January 1956 to August 1957 are presented in Table 1, in Portuguese. The experiment consisted of 3 replications of the treatments: without fertilizer and with fertilizer (40 Kg of N, from ammonium sulfate; 100 Kg of P(2)0(5) from superphosphate and 40 Kg K2 O, from potassium chloride). Four complete stools (stalks and leaves) were harvested from each treatment, and the plants separated in stalks and leaves, weighed, dried and analysed every month from 6 up to 15 months of age. The data obtained for fresh and dry matter production are presented in table 2, and in figure land 2, in Portuguese. The curves for fresh and dry matter production showed that fertilized and no fertilized sugar cane with 6 months of age presents only 5% of its total weight at 15 months of age. The most intense period of growth in this experiment is located, between 8 and 12 months of age, that is between December 1956 and April 1957. The dry matter production of sugar cane with 8 and 12 months of age was, respectively, 12,5% and 87,5% of the total weight at 15 months of age. The growth of sugar cane in relation to its age follows a sigmoid curve, according to the figures 1, 2 and 3. The increase of dry matter production promoted by using fertilizer was 62,5% when sugar cane was 15 months of age. The concentration of the elements (tables 4 and 5 in Portuguese) present a general trend of decreasing as the cane grows older. In the stalks this is true for all elements studied in this experiment. But in the leaves, somme elements, like sulfur and silicon, appears to increase with the increasing of age. Others, like calcium and magnesium do not show large variations, and finally a third group, formed by nitrogen, phosphorus and potassium seems to decrease at the beginning and later presents a light increasing. The concentration of the elements was higher in the leaves than in the stalks from 6 up to 15 months of age. There were some exceptions. Potassium, magnesium and sulfur were higher in the stalks than in the leaves from 6 up to 8 or 9 months of age. After 9 months, the leaves presented more potassium, magnesium and sulfur than the stalks. The percentage of nitrogen in the leaves was lower in the plants that received fertilizer than in the plants without fertilizer with 6, 7, 8, 10, 11 and 13 months of age. This can be explained by "dilution effect". The uptake of elements by 4 stools (stalks and leaves) of sugar cane according to the plant age is showed in table 6, in Portuguese. The absorption of all studied elements, nitrogen, phosphorus, potassium, calcium, magnesium, sulfur and silicon, was higher in plants that received fertilizer. The trend of uptake of nitrogen and potassium is similar to the trend of production of dry matter, that is, the maximum absorption of those two nutrients occurs between 9 and 13 months of age. Finaly, the maxima amounts of elements absorbed by 4 stools (stalks and leaves) of sugar cane plants that received fertilizer are condensed in the following table: Element Maximum absorption in grams Age of the plants in months Nitrogen (N) 81.0 14 Phosphorus (P) 6.8 15 Potassium (K) 81.5 15 Calcium (Ca) 19.2 15 Magnesium (Mg) 13.9 13 Sulfur (S) 9.3 15 Silicon (Si) 61.8 15 It is very interesting to note the low absorption of phosphorus even with 100 kg of P2O5 per hectare, aplied as superphosphate. The uptake of phosphorus was lower than calcium, magnesium and sulfur. Also, it is noteworthy the large amount of silicon absorbed by sugar cane.