Accessibility to primary health care in low-density regions. A case study: NUTS III - Baixo Alentejo - Portugal / A acessibilidade aos cuidados de saúde primários em regiões de baixa densidade. Caso de estudo: NUTS III - Baixo Alentejo - Portugal

Abstract This study diagnosed the situation regarding the physical accessibility of the resident population to primary health care, based on the characteristics of the population served, their spatial distribution in the territory, based on space-time analysis. Thus, bearing the different means of transport available and the specific features of a low-density territory, we considered several mobility profiles under analysis, and selected the Baixo Alentejo as the study area. In methodological terms, besides using the location of primary health facilities and their areas of influence, the use of the road network and its restrictions, we selected the use the new 1x1 km grid, recently implemented throughout the EU (European Union), instead of using the statistical units or administrative boundaries. Its advantages allow overcoming some of the issues of the usual base cartography. The final results can be divided into two groups: conclusions related to the methodologies used and conclusions related to the accessibility of primary health care equipment in the study area.

Resumo Decidiu-se realizar o diagnóstico de situação sobre a acessibilidade física da população residente aos cuidados de saúde primários, baseada nas características da população servida, na sua distribuição espacial, tendo por base uma análise espaço/tempo, permitindo avaliar essa mesma acessibilidade. Deste modo, e tendo em consideração os diversos modos de transporte disponíveis, bem como as características específicas de um território de baixa densidade, foram considerados diversos perfis de mobilidade, sendo a área de estudo escolhida o Baixo Alentejo. Em termos metodológicos, além da utilização da localização dos equipamentos de saúde primários e das suas áreas de influência, da utilização da rede viária e das suas restrições, optou-se pela utilização de uma cartografia base menos convencional. Em vez das unidades estatísticas associadas às unidades administrativas, optou-se pela utilização da nova quadrícula de 1x1 km, implementada recentemente em toda a União Europeia (UE). A utilização desta quadrícula permite ultrapassar alguns dos problemas apresentados pela cartografia base habitual. Os resultados podem-se dividir em dois grupos: conclusões relacionadas com as metodologias utilizadas, e conclusões relacionadas com a acessibilidade aos equipamentos de saúde primários na área de estudo.

MicroRNA-152 and microRNA-448 inhibit proliferation of colorectal cancer cells by targeting Rictor / 南方医科大学学报

OBJECTIVE@#To screen the microRNAs (miRNAs) targeting Rictor and investigate their effects in regulating the biological behaviors of colorectal cancer (CRC).@*METHODS@#Human colorectal cancer cell line KM12SM was transfected with the miRNAs targeting Rictor identified by prediction software to test inhibitory effects of these miRNAs on Rictor expression using qRT-PCR and Western blotting. Dual luciferase reporter assay was used to further confirm the binding of these miRNAs to the 3'UTR of Rictor mRNA. Cell survival and colony formation assays were used to investigate the effects of these miRNAs on survival and colony formation in KM12SM cells.@*RESULTS@#miR-152 and miR-448 were identified as the Rictor-targeting miRNAs, which significantly inhibited the expression of Rictor in KM12SM cells ( < 0.05). The two miRNAs were confirmed to bind to the 3'UTR of Rictor mRNA and significantly inhibited luciferase activity in KM12SM cells ( < 0.01, < 0.05); they also showed activities of posttranscriptional modulation of Rictor. Overexpression of miR-152 and miR-448 both significantly inhibited the growth and colony formation of KM12SM cells.@*CONCLUSIONS@#miR-152 and miR-448 can down-regulate the protein expression of Rictor by targeting Rictor mRNA to negatively regulate the growth and colony formation of colorectal cancer cells.

Effects of different fluid replenishment methods on internal environment, body thermal regulation response and severe heatstroke of 5-km armed cross-country training soldiers / 中华危重病急救医学

To explore the effects of different fluid replenishment methods on the internal environment, body thermal regulatory response and severe heatstroke of 5-km armed cross-country training soldiers. Methods A Special Force officers and soldiers who participated in 5-km armed cross-country training (2-3 times a week, 25-30 minutes each time for 3 weeks) during summer training from June to July in 2018 were enrolled, and they were divided into three groups according to the random number table, with 300 trainees in each group. 200 mL of drinking fluids were given to each group 15 minutes before and after each 5-km armed cross-country training: A group with boiled water, B group with purified water, and C group with beverage prepared by pharmaceutical laboratory of the 990th Hospital of PLA Joint Logistics Support Force (100 mL containing 6 g carbohydrates, 42 mg sodium, and 11 mg potassium). The venous blood was collected before and after the last training or during the onset of severe heatstroke to do the following tests: serum cardiac troponin I (cTnI, chemiluminescence), MB isoenzyme of creatine kinase (CK-MB, immunosuppressive), serum creatinine (SCr, enzymatic method), urea nitrogen (BUN, enzymatic method), alanine aminotransferase (ALT, tryptase), aspartate transaminase (AST, tryptase), and Na+, K+, Cl- (electrode method). The heart rate (HR) and core temperature (Tc, anal temperature) were monitored at the same time. The amount of sweat in training and the occurrence of severe heatstroke were also recorded. Results There was no significant difference in heart, liver, kidney function, electrolyte and body heat regulation reaction among three groups of 5-km armed cross-country trainees before training. Compared with before training, the levels of serum cTnI, CK-MB, SCr, BUN, ALT, AST, HR and Tc were significantly increased after training or during the onset of severe heatstroke in three groups, while the contents of Na+, K+, Cl- were significantly decreased, but the increase or decrease of group C was relatively smaller compared with group A and group B [cTnI (μg/L): 0.9 (0.6, 1.4) vs. 1.1 (0.7, 2.8), 1.0 (0.6, 3.3); CK-MB (U/L): 7.0 (5.0, 11.0) vs. 9.0 (6.0, 14.5), 8.0 (6.0, 15.0); SCr (μmol/L): 92.09±18.64 vs. 102.78±18.77, 103.64±20.07; BUN (mmol/L): 7 (6, 9) vs. 9 (8, 11), 10 (8, 13); ALT (U/L): 27 (22, 34) vs. 36 (30, 43), 34 (27, 43); AST (U/L): 37 (31, 48) vs. 41 (34, 50), 39 (34, 51); HR (bpm):87.01±17.07 vs. 95.88±21.06, 96.59±22.04; Tc (℃): 37.73±0.81 vs. 38.03±1.05, 38.10±1.04; Na+ (mmol/L):150.14±3.86 vs. 144.18±8.89, 144.04±9.39; K+ (mmol/L): 4.32±0.57 vs. 4.15±0.62, 4.13±0.51; Cl- (mmol/L):100.43±3.71 vs. 98.42±4.24, 98.41±4.58; all P < 0.01]. The incidence of severe heatstroke in group C was significantly lower than that in group A and group B [1.67% (5/300) vs. 5.00% (15/300), 5.33% (16/300), χ2 = 6.424, P = 0.040]. There was no significant difference in sweating volume in groups A, B, C (g: 370.47±48.71, 370.85±50.66, 370.17±50.21, F = 0.014, P = 0.986). There was no significant difference in the above indexes between group A and group B (all P > 0.05). Bi-classification Logistic regression analysis showed that the increase of HR, Tc and excessive loss of Na+, K+, Cl- were risk factors for severe heatstroke [odds ratio (OR) was 0.848, 0.138, 1.565, 17.996 and 2.328 respectively, all P < 0.01]. Conclusions Timely supplementation of carbohydrate, sodium and potassium ions can effectively change the internal environment and body heat regulation reaction of 5-km armed cross-country trainees, so as to reduce the occurrence of severe heatstroke. The increases of HR, Tc and excessive loss of Na+, K+, Cl- are risk factors for severe heatstroke.

Effects of training burnout and sleep quality on heat regulation response and severe heatstroke in people performed 5-km armed cross-country training / 中华危重病急救医学

Objective To explore the relationship between training burnout, sleep quality and heat regulation response, severe heatstroke in people performed 5-km armed cross-country training. Methods 600 male officers and soldiers who participated in 5-km armed cross-country training in summer from 2017 to 2018 were enrolled. All trainees participated in 5-km armed cross-country training in environment with ambient temperature > 32 ℃ and (or) humidity > 65%. They were divided into two groups according to whether severe heatstroke occurred during 5-km armed cross-country training. The age, military age, body mass index (BMI), physical fitness score, external environment (such as ambient temperature, relative humidity, wind speed, heat index), training burnout score and Pittsburgh sleep quality index scale (PSQI) score, heart rate (HR), core temperature (Tc), sweating volume and serum Na+, K+, Cl- levels were compared between the groups. The risk factors of severe heatstroke during 5-km armed cross-country training were screened by binary multivariate Logistic regression analysis. Results There were 26 cases of severe heatstroke in 600 trainees who participated in 5-km armed cross-country training, with an incidence of 4.33%. There was no significant difference in age, military age, BMI, physical fitness score and external environment of 5-km armed cross-country training between people with or without severe heatstroke. Compared with those without severe heatstroke, the dimensions of training burnout and the total average scores of training burnout of severe heatstroke personnel before 5-km armed cross-country training were increased significantly (physical and mental exhaustion score: 12.4±2.5 vs. 9.4±3.5, training alienation score: 8.8±2.8 vs. 5.8±2.3, low sense of achievement score: 8.2±2.7 vs. 5.6±2.3, total score of training burnout: 9.8±3.2 vs. 6.9±3.2, all P < 0.01), all factors except daytime dysfunction (DD) of PSQI and total PSQI score were also increased significantly [sleep quality (SQ) score: 1.0 (1.0, 2.0) vs. 1.0 (1.0, 1.0), fall asleep time (SL) score: 2.0 (1.0, 3.0) vs. 1.0 (1.0, 1.0), sleep time (SH) score: 1.0 (0.8, 2.0) vs. 1.0 (0, 1.0), sleep efficiency (SE) score: 1.0 (0, 1.0) vs. 0 (0, 0.8), sleep disorder (SD) score: 2.0 (1.0, 3.0) vs. 1.0 (0, 2.0), total PSQI score: 1.0 (1.0, 2.0) vs. 1.0 (0, 1.0), all P < 0.01], HR was increased significantly at onset (bpm: 120.00±10.57 vs. 86.49±14.91, P < 0.01), Tc was increased significantly (℃: 41.46±0.57 vs. 37.97±0.83, P < 0.01), serum electrolyte contents were decreased significantly [Na+ (mmol/L): 130.54±5.97 vs. 143.15±10.56, K+ (mmol/L): 3.72±0.44 vs. 4.37±0.50, Cl- (mmol/L):97.58±4.80 vs. 102.10±2.39, all P < 0.01], and the amount of sweat during training was increased significantly (g: 395.81±16.16 vs. 371.88±40.76, P < 0.01). Binary multivariate Logistic regression analysis showed that total score of training burnout [odd ratio (OR) = 0.653, 95% confidence interval (95%CI) = 0.563-0.757], total PSQI score (OR =0.693, 95%CI = 0.525-0.916), HR (OR = 0.871, 95%CI = 0.838-0.908), Tc (OR = 0.088, 95%CI = 0.043-0.179), sweating volume (OR = 0.988, 95%CI = 0.979-0.997), Na+ (OR = 1.112, 95%CI = 1.069-1.158), K+ (OR = 13.900, 95%CI = 5.343-36.166), Cl- (OR = 1.393, 95%CI = 1.252-1.550) were independent risk factors for severe heatstroke during 5-km armed cross-country training (all P < 0.01). Conclusion Increase in training burnout, total PSQI score, excessive changes of body heat regulation response and excessive loss of Na+, K+, Cl- in serum are independent risk factors for severe heatstroke during 5-km armed cross-country training under the same conditions with high temperature and humidity environment.

Relationship between physiological parameters changes and severe heatstroke induced by 5-km armed cross-country training / 中华危重病急救医学

Objective@#To explore the relationship between physiological parameters changes and severe heatstroke induced by 5-km armed cross-country training.@*Methods@#A total of 521 male officers and soldiers from a special team who participated in the summer training of 5-km armed cross-country training from year 2016 to 2017 were enrolled. All trainees participated in 5-km armed cross-country training in high temperature and humidity environment of ambient temperature > 32 ℃and (or) relative humidity > 65%. The trainees were divided into two groups according to the incidence of severe heatstroke in the course of training. The age, enlistment time, constitution score, body mass index (BMI), external environment (ambient temperature, relative humidity, wind speed, heat index) of trainees of the two groups, and the change rates of arterial blood oxygen saturation (SaO2), body temperature, pulse and blood pressure within 5 minutes after the 5-km armed cross-country training were compared between the two groups. The risk factors of severe heatstroke were screened by two classified Logistic regression analysis, and the predictive value of various risk factors of severe heatstroke was analyzed by the receiver operator characteristic curve (ROC).@*Results@#In 521 trainees of 5-km armed cross-country training, 29 trainees suffered from severe heatstroke accounting for 5.57%. There was no significant difference in the age, enlistment time, constitution score, BMI, or external environment during 5-km armed cross-country training between severe heatstroke group and non-severe heatstroke group. Compared with those without severe heatstroke, the descending rates of body temperature, pulse, blood pressure and SaO2 increased rate within 5 minutes after 5-km armed cross-country training of severe heatstroke trainees were significantly decreased [temperature descending rate: (0.67±0.30)% vs. (1.43±1.28)%, pulse descending rate: (7.53±5.21)% vs. (13.48±8.07)%, blood pressure descending rate: (9.28±6.84)% vs. (19.42±7.73)%, SaO2 increased rate: (0.51±0.39)% vs. (1.50±1.43)%, all P < 0.01]. Two classification Logistic regression analysis showed that the temperature descending rate [odds ratio (OR) = 0.485, 95% confidence interval (95%CI) = 0.289-0.817], pulse descending rate (OR = 0.903, 95%CI = 0.845-0.965), blood pressure descending rate (OR = 0.841, 95%CI = 0.790-0.896), and SaO2 increased rate (OR = 0.421, 95%CI = 0.250-0.711) were the risk factors for severe heatstroke during 5-km armed cross-country training (all P < 0.01). ROC curve analysis showed that temperature descending rate [area under ROC curve (AUC) = 0.659, 95%CI = 0.604-0.714], pulse descending rate (AUC = 0.730, 95%CI = 0.762-0.900), blood pressure descending rate (AUC = 0.831, 95%CI = 0.659-0.801), SaO2 increased rate (AUC = 0.711, 95%CI = 0.655-0.767) could be used for the incidence of severe heatstroke prediction during 5-km armed cross-country training (all P < 0.01), and the predicted value was the same.@*Conclusions@#Under the same conditions, the severe heatstroke during 5-km cross-country training is closely related to the descending rates of body temperature, pulse, and blood pressure as well as SaO2 increased rate within 5 minutes after the training, whose predictive values for severe heatstroke were the same.

Inductive effect of BuShenYangGuTang on apoptosis of multiple myeloma cell line KM3 and its mechanism / 天津医药

Objective To study the effects and mechanisms of BuShenYangGuTang on proliferation and apoptosis of multiple myeloma cell line KM3. Methods The inhibitory effect of BuShenYangGuTang on cell proliferation was assessed by CCK-8. BuShenYangGuTang induced KM3 cell cycle arrest was analyzed by flow cytometry after propidium iodide staining. Flow cytometry was also used to analyze the cell apoptosis after Annexin V-FITC staining. The expressions of Bcl-2, Bax and NF-κB were determined by RT-qPCR and Western blot assay. Results BuShenYangGuTang inhibited the KM3 cell proliferation in a dose-dependent manner. The expression levels of Bcl-2 and NF-κB were decreased, the expression level of Bax was increased, and the cell cycle was arrested in G0 / G1 phase after treatment with BuShenYangGuTang. Conclusion BuShenYangGuTang could inhibit the proliferation, arrest cell cycle and induce the apoptosis in KM3 cells, which may be related to the abnormal expressions of Bcl-2, Bax and NF-κB.

Chemical constituents and pharmacological activities of Ferula sinkiangensis K.M. Shen:research advances / 国际药学研究杂志

Ferula sinkiangensis K.M. Shen is a kind of endangered national plant medicine in the northern of Xinjiang,China. It mainly contains sesquiterpene coumarins,sulfides,polysaccharides,aromatic compounds and other effective chemical constitu-tions;Chromatography is used to build the fingerprint and to control the medicine quality in most cases. Both raw plant medicine and specific compounds possess pharmacologic activities such as antibacterial,insecticidal,anti-allergic,and antitumor properties. The present paper summarizes the resource distribution,chemical constituents,fingerprints and pharmacological activities of F. sinkian-gensis,to provide reference for its systematic research,effective quality control and efficient development and utilization.

Induction and enhancement of the novel marine-derived Alternaria tenuissima KM651985 laccase enzyme using response surface methodology: Application to Azo and Triphenylmethane dyes decolorization.

In our search for a laccase producer from unconventional sources, the marine derived fungus Alternaria tenuissima KM651985 was isolated from decayed wood. It was identified on the basis of morphological and molecular taxonomy and got the Genbank accession number KM651985. Two statistical experimental designs were employed to enhance laccase production. At first, a two level Plackett-Burman design was employed to screen the medium constituents and inducers that significantly affect the enzyme production. Second experiment was important for optimization of the most effective constituents and inducers using central composite design. Applying the above methods revealed that guaiacol (2 mM), copper sulphate (3 mM) and wheat bran (46.82 g/l) were the most effective factors affecting laccase production. A maximal enzyme activity of 91.84 U/ml was more than 6.33-folds the activity obtained using the basal medium. Application of A. tenuissima KM651985 culture medium containing laccase to decolorize two structurally different synthetic dyes was done successfully. This is the first report on the statistical optimization of the marine-derived A. tenuissima KM651985 laccase enzyme and its applications for dyes decolorization.

Establishment and comparison of obese asthma models in different strains of mice / 中国药理学通报

Aim To establish and compare asthma models among different strains of obese mice. Methods Different strains of SPF female mice, namely Kunming ( KM ) , C57BL/6J and BALB/c mice, were randomly divided into four groups ( control group, asthma group, obesity group and obese asthma group). The mice were fed a high-fat diet or a normal diet for 12 weeks, following which they were sensitized and challenged with ovalbu-min ( OVA) or phosphate-buffered saline ( PBS) . Body weight, fat weight, liver weight, Lee′s index, OVA-specific IgE concen-tration in bronchoalveolar lavage fluid ( BALF) , serum total cho-lesterol ( TC) and triglyceride ( TG) levels, and lung and adi-pose morphologies were evaluated. The specific airway resistance ( sRaw) was measured using double-chamber plethysmography. Results The mice on a high-fat diet showed a more rapid in-crease in body weight compared with those on a normal diet. Af-ter 12 weeks of feeding, body, fat, and liver weights and Lee′s index were higher for the obese mice than for the lean mice. The adipocyte cross-sectional area was significantly greater in the obese BALB/c and KM mice than in their lean counterparts;the C57BL/6J groups showed no significant differences. The BALB/c mice demonstrated more significant symptoms of acute asthma, local inflammation and airway hyper-responsiveness ( AHR ) . Conclusion Compared with C57BL/6J and KM mice, BALB/c mice fed a high-fat diet and sensitized and challenged with OVA provide the most suitable model for evaluating the relationship between obesity and asthma.

Construction of type 1 diabetic KM mice model induced by single high dose and multiple low dose of streptozotocin / 中国生化药物杂志

Objective To explore an optimal method of producing STZ-induced type 1 diabetic KM mice model by investigating the molded rate of single high dose and multiple low dose of STZ injection.Methods Sixty KM mice were randomly divided into 4 groups(n=15), two control groups and two model groups.In the two control groups, one was blank control and the other was negative control.Mice in the blank control group treated with no injection, but mice in the negative control group were treated with injection of citric acid salt buffer.For the two model groups, one was single high-dose group, 150 mg/kg STZ was injected only once.The other was multiple low-dose group, 50 mg/kg STZ was injected for 5 consecutive days.After the last injection, daily food and water intake were tested, blood glucose level and body weight were examined once a week for 4 consecutive weeks. Results Mice in the two model groups showed typical features of diabetes.The blood glucose levels in the two model groups were significantly higher than in the two control groups ( P <0.05 ) .For the two model groups, the molded rate of 150 mg/kg and 50 mg/kg group were 60% and 53.33%respectively at 1st week, but at the 4th week, they were 60% and 80% respectively.The mortality of these two groups at the 4th week was 33.33% and 20% respectively.Moreover, the blood glucose level in multiple low-dose group increased stably from the 2nd week to the 4th week.Conclusion The multiple low-dose STZ injection (50 mg/kg for 5 consecutive days) is an optimal method for producing KM mice model of type 1 diabetes mellitus.

Rapidly detecting the sperm concentration in mice and rats based on enzyme-labeled instrument / 中国生化药物杂志

Objective To establish a rapid and accurate method, and to determine the density of mice and rats sperm with enzyme-labeled instrument.Methods ①The optimal wavelengths and the regression equation set up: After six Kunming mice and six Sprague-Dawley rats were sacrificed, the left epididymis was separated and fully cut up in phosphate buffer saline.With water bath, the sperm were fully dissociated.Using the enzyme-labeled instrument to detect the wavelength absorbance respectively under different wavelength and fitting absorbance curve.The best wavelength will be the most close to 1 of the correlation coefficient ( R2 ) and the standard deviation of minimum.After ten Kunming mice and ten Sprague-Dawley rats were sacrificed,the sperm suspension of different concentration gradient were got.The regression equation of the sperm density and absorbance was established by using enzyme-labeled instrument and haemocytometer.②The test of sperm absorbance stability: Mice and rats,six respectively,were used to make the sperm suspension.Samples were put in room temperature (25℃) or 37℃water bath continued,and after water bath about 0,20,30,40,50, 60 min,the change of absorbance was recorded.③The regression equation verification:The mice were administrated orally with 20% ethanol solution for 30 days to make oligospermia.In order to verify the new method, two different method were used to get the sample sperm.Results The optimal absorbancy-sperm density curve could be established at 380 nm.The means of KM mice sperm count ( x1 ) and absorbance ( y1 ) are showed to be the linear function,and the linear regression equation is y1 =2 ×10 -9 x1 +0.0648, R2 =0.9743.The means of SD rat sperm count ( x2 ) and absorbance (y2) are showed to be the linear function,and the linear regression equation is y2 =5 ×10 -9x2 +0.0621,R2 =0.9940.SD rat sperm suspension liquid after 60 min in water bath, absorbance value at 0 min significantly decreased(P<0.05), but at room temperature after 40 min significantly increased ( P<0.05); KM mice sperm suspension in the water bath and under the condition of normal temperature after 50 min, compared with the 0 min, absorbance value increased significantly(P<0.05).Compared with control group, sperm density of ethanol oligozoospermia group by enzyme standard detector and standard curve calculation were significantly decreased ( P <0.05 );compared with absorbancy-sperm density equation, determination of ethanol oligozoospermia group of sperm density by cell counting plate method had not significant difference.The results suggested absorbancy-sperm density equation could effectively detect the reduction of the mice sperm in oligospermia.Conclusion Using enzyme-labeled instrument to set up the curve of absorbancy-sperm density equation can estimate the sperm density of mice and rats rapidly and exactly.

Production and Partial Characterization of Pectinase from Snail (Archachatina marginata) Gut Isolates.

Aim: To identify isolates for industrial production of pectinase. Methodology: The isolates were screened for pectinolytic activity using pectin as substrate. Enzyme activity was expressed as mg of glucose equivalent released per ml of crude enzyme solution per second. The kinetic parameters of the enzyme were determined to obtain the optimum pH, temperature, Km and Vmax. Pectinase produced from Bacillus subtillis was immobilized on chitosan and its activity was compared with that of free enzyme. Results: Pectinase from Bacillus subtilis was screened to have the highest enzymatic activity among bacterial isolates while pectinase from Aspergillus niger had the highest enzymatic activity among fungal isolates. High yield of pectinase enzyme was obtained from B. subtilis after 24hrs with activity of 4.01×10-4 mg/ml/sec while high yield of pectinase was obtained from A. niger on the 5th day with activity of 2.07×10-4 mg/ml/sec. The optimum pH for pectinase produced from B. subtilis and A. niger were 8 and 6, respectively. The optimum temperature for pectinase from B. subtilis and A. niger were at 50ºC and 40ºC, respectively. The Vmax and Km of pectinase from B. subtilis and A. niger were 16.88×10-4 (mg/ml/sec) and 10 (mg/ml); 9.29×10-4 (mg/ml/sec) and 30 (mg/ml), respectively. Optimum temperature and pH of immobilized pectinase were 70ºC and 4.0, respectively. Residual activity of immobilized enzyme was 92% after storage at 4ºC for 14 days. Conclusion: This study revealed that Bacillus subtilis from snail gut may be considered as a good candidate for industrial production of pectinase.

Purification and Characterization of α-Glucan Phosphorylase Isoform Pho 2 from Spinach Leaves.

α-Glucan phosphorylase is an important enzyme of carbohydrate metabolism. In spinach leaves, it has been reported in two multiple forms viz. Pho 2 (cytosolic) and Pho 1 (plastidial). Here, we extracted and purified Pho 2 form of α-glucan phosphorylase from spinach using salting out with ammonium sulfate, desalting using Sephadex G-25 chromatography, anion exchange chromatography using DEAE-Sepharose and gel filtration chromatography using Sepharose-4-B. The purified enzyme had a specific activity, 150 units/mg protein. There was 38% recovery and 652 fold purification after final Sepharose-4B chromatography. The purified enzyme showed a single protein band on SDS sodium dodecyl sulfate polyacrylamide gel electrophoresis having molecular weight 94,000±2000 daltons. The native molecular weight is found to be 188,000±3000 daltons as determined using gel filtration chromatography over Sephadex G-200. The Pho2 exhibited optimum pH at pH 6.0 with two half pH optima at pH 5.2 and pH 7.0. The optimum temperature of Pho2 is found to be 37ºC with two half temperature optima at 30ºC and 40ºC. The Km value of the enzyme for starch and glucose-1-phosphate is found to be 116 μg/mL and 0.55 mM, respectively.

How to Choose Drug Dosage for Human Experiments Based on Drug Dose Used on Animal Experiments: A Review.

The orthodontic tooth movement is a biological response to orthodontic force. The biological response is very strongly-related to local bone metabolism. There is a strong evidence in the literature that bone metabolism can be altered by drugs. There are various studies published in dental journals on administration of drugs for the purpose of affecting orthodontic tooth movement both for augmentation of anchorage and to increase the rate of tooth movement. Most of these studies are animal studies. The aim of this article is to give insight to how to convert drug dose from animal studies to human trails. Dose per kilogram of body weight for one species is not the same for another species, it has to be converted first based on body surface area (BSA)normalization method. BSA correlates well across several mammalian species with several parameters of biology, including oxygen utilization, caloric expenditure, basal metabolism, blood volume, circulating plasma proteins, and renal function.

Ripening and shelf life of 'BRS Caipira' banana fruit stored under room temperature or refrigeration / Amadurecimento e vida útil de frutos de bananeira 'BRS Caipira' armazenados em temperatura ambiente ou refrigeração

BRS Caipira variety, internationally known as 'Yangambi km 5', is an alternative to meet the demand of 'Maçã'-type fruit due to its resistance to Panama disease. This study had the objective of generating information about 'BRS Caipira' fruit ripening and cold storage potential. For the ripening study fruits were stored under room temperature conditions (25±2°C / 58±6% U.R.) and assessed for postharvest life evaluation and characterization of seven maturity stages based on peel color: completely green - MS1; green with yellow traces - MS2; more green than yellow - MS3; more yellow than green - MS4; yellow with green tips - MS5; completely yellow - MS6; yellow with brown spots - MS7. For the cold storage potential study, fruits at MS1 were cold stored (14±1°C / 53±2% U.R.) for 28 days. Weekly, fruits were transferred to room temperature to ripen until MS6 when were assessed for quality attributes. Ripening of 'BRS Caipira' fruit was characterized as slow between MS1 and MS2 (averaging five days), then fast between MS2 and MS6 (up to four days in average), and undergoing determinant changes between MS6 and MS7: pulp yield reached 80%, titratable acidity reduced by 50% and ratio increased by 78%. Cold storage extended shelf life by up to 19 days as compared with control, without visible symptoms of chilling injury, although tends to reduce soluble solids in ripe fruit. Maximum recommended time for storage of 'BRS Caipira' fruit at 14°C is 21 days, since it allows a few more days under room temperature until fruit reach MS6.

A variedade BRS Caipira, internacionalmente conhecida como 'Yangambi km 5', é uma alternativa para atender a demanda de frutos do tipo Maçã, devido à sua resistência ao mal-do-Panamá. Este estudo objetivou gerar informações sobre o amadurecimento e o potencial de armazenamento refrigerado de frutos de 'BRS Caipira'. Para o estudo do amadurecimento, frutos foram armazenados sob temperatura ambiente (25±2°C / 58±6% U.R.) e avaliados para vida pós-colheita e caracterização de sete estádios de maturação baseados na cor da casca: completamente verde - MS1; verde com traços amarelos - MS2; mais verde que amarelo - MS3; mais amarelo que verde - MS4; amarelo com pontas verdes - MS5; completamente amarelo - MS6; amarelo com manchas marrons - MS7. Para o estudo de potencial de armazenamento refrigerado, frutos em MS1 foram armazenados em câmara fria (14±1°C / 53±2% U.R.) por 28 dias. Semanalmente, uma amostra de frutos era retirada para a temperatura ambiente e avaliados para atributos de qualidade. O amadurecimento de frutos de 'BRS Caipira' foi caracterizado como lento entre MS1 e MS2 (cinco dias em média), rápido entre MS2 e MS6 (até quatro dias em média), e que passa por alterações determinantes entre MS6 e MS7: rendimento em polpa atingiu 80%; acidez reduziu pela metade; e ratio aumentou 78%. A refrigeração prolongou a vida útil dos frutos em até 19 dias em relação ao controle, sem sintomas visíveis de danos por frio, embora tenda a reduzir o teor de sólidos solúveis nos frutos maduros. O período máximo recomendado para armazenagem de frutos de 'BRS Caipira' a 14°C é de 21 dias, tempo que permitiria mais alguns dias em temperatura ambiente para os frutos atingirem o estádio MS6.

Phenylpropanoid enzymes, phenolic polymers and metabolites as chemical defenses to infection of Pratylenchus coffeae in roots of resistant and susceptible bananas (Musa spp.).

Activity differences of the first (phenylalanine ammonia lyase, PAL) and the last (cinnamyl alcohol dehydrogenase, CAD) enzymes of phenylpropanoid pathway in the roots of resistant (Yangambi Km5 and Anaikomban) and susceptible (Nendran and Robusta) banana cultivars caused by root lesion nematode, Pratylenchus coffeae, were investigated. Also, the accumulation of phenolics and deposition of lignin polymers in cell walls in relation to resistance of the banana cultivars to the nematode were analyzed. Compared to the susceptible cultivars, the resistant cultivars had constitutively significantly higher PAL activity and total soluble and cell wall-bound phenolics than in susceptible cultivars. The resistant cultivars responded strongly to the infection of the nematode by induction of several-time higher PAL and CAD enzymes activities, soluble and wall-bound phenolics and enrichment of lignin polymers in cell wall and these biochemical parameters reached maximum at 7th day postinoculation. In addition, profiles of phenolic acid metabolites in roots of Yangambi Km5 and Nendran were analyzed by HPLC to ascertain the underlying biochemical mechanism of bananas resistance to the nematode. Identification and quantification of soluble and cell wall-bound phenolic acids showed six metabolites and only quantitative, no qualitative, differences occurred between the resistant and susceptible cvs. and between constitutive and induced contents. A very prominent increase of p-coumaric, ferulic and sinapic acids, which are precursors of monolignols of lignin, in resistant cv. was found. These constitutive and induced biochemical alterations are definitely the chemical defenses of resistant cvs. to the nematode infection.

Biochemical studies in relation to glutamate dehydrogenase in piper nigrum (L.).

Black pepper, the king of spices, the most important and widely used spice in the world is the dried mature berries of P. nigrum L. Inorganic nitrogen is assimilated into amino acids glutamine, glutamate, asparagines and aspartate which serve as important nitrogen carriers in plants. The enzymes glutamine synthetase (GS) EC. 6.3.1.2, glutamate synthase (GOGAT) E.C. 1.4.1.14, glutamate dehydrogenase (GDH) E.C.1.4.1.4, aspartate aminotransferase and asparagine synthetase are responsible for biosynthesis of these amino acids. Although extensive studies have been carried out on the epidemiology of the diseases and agronomical aspects of P. nigrum, very few studies have been made on the enzymological aspects especially that associated with the nitrogen assimilation. A better understanding of the various properties of the enzyme will be valuable in the future studies concerned with Nitrogen metabolism in P. nigrum. Main objective of present study was to standardize the extraction and study the properties of GDH from P. nigrum L. Effect of additives and its optimum concentration were determined, standardized homogenizing medium was formulated, standardized assay system formulated, different properties like pH optimum, Substrate saturation and km value determined; it’s all in agreement with the previously reported values, result indicate NADP+/NADPH is the better substrate i.e. the enzyme activity is contributed by chloroplastic form of GDH than by the mitochondrial form.

Population genetic quality analysis of outbred KM mice from Shanghai Seed Center / 中国比较医学杂志

Objective To analyze and evaluate the population genetic quality of outbred KM mice from National Rodent Seed Center ( Shanghai ) .Methods A total of 30 outbred KM mice were randomly chosen.The genetic characteristics of the population were determined by PCR and STR scanning using 30 selected microsatellite loci. Popgen1.32 software was used to process the data.Results Thirty microsatellite loci shared 123 alleles in the KM mouse population.The average effective allele number and the average heterozygosity were 2.3989 and 0.5342, respectively.The average polymorphism information content ( PIC ) was 0.4735.Conclusions The outbred KM mouse population of Shanghai Seed Center has genetic stability and genetic diversity, and is satisfied with the genetic characteristics of closed colony laboratory animal.

Isolation and Identification of Xylanolytic Enzyme from an Effective Strain Bacillus Licheniformis Isolated from the Decaying Wood.

A new species of Bacillus licheniformis produced extracellular xylanase under submerged fermentation when wheat bran is used as carbon source. The xylan is the most common hemicellulosic polysaccharide in food industry and agricultural wastes, comprising a backbone of xylose residues linked by β-1,4 glycosidic bonds. Bacillus licheniformis has been shown to be a promising organism for enhanced production of xylanases & β-xylosidase under submerged fermentation (SmF). The optimization of cultural conditions and carbon, nitrogen sources for enzymes production. The bacterial strain Bacillus licheniformis was cultivated using as substrate xylan, wheat bran, corn straw, corncob, and sugarcane bagasse. Wheat bran has been a good xylanase (16.8U/ml) & β xylosidase (5.6U/ml) activity after 48h of fermentation. Maximum enzyme activity was observed in xylan as carbon source and peptone as nitrogen source. Both crude enzymes were characterized and a bacterial xylanase shows optimum pH for xylanase activity at 6.5 & β xylosidase were found to be 6.0. The optimum temperatures were 450C for both and they were thermally stable up to 500C. The parameters of Vmax and Km obtained using Line weaver-Burk plot method were 277.7μmol / min/mg and 5.26 mg /L correspondingly.

Formulation and evaluation of Norfloxacin gastro retentive drug delivery systems using natural polymers.

Gastric retentive floating drug delivery system (GFDDS) is enabled the prolonged continuous input of the drug to the upper parts of the gastrointestinal (GI) tract and improves the bioavailability of medications with narrow absorption window. The design of the delivery system is based on the controlled release formulation with floating and swelling features in order to prolong the gastric retention time of the drug delivery systems. In the present study norfloxacin as candidate, guar gum, sodium CMC, HPMC15 KM is studied along with other excipients like PVP K30 (binder), sodium bicarbonate microcrystalline cellulose were used in different concentrations to get the desired controlled release profile over a period of 12 hrs. All the formulations were evaluated for buoyancy lag time, duration of buoyancy, dimensional stability, drug content and in vitro drug release profile. Based on the in vitro studies carried out for the optimized formulation by dissolution the performance of the developed formulation promises to be efficient in controlling the drug release rate with the guar gum , a natural polymer.