Refractometry as an alternative to the biuret method for measuring total serum proteins in Podocnemis expansa (Podocnemididae) and Phrynops geoffroanus (Chelidae)

Pessoa, Lourdes Marina Bezerra; Lima, Maíra Gonçalves da Mota; Carneiro, Filipe T.; Zanani, Nathalia Salgado; Silva, Thamiris Figueiredo; Lima, Mariana Accioly; Abrahim, M. A.; Paludo, Giane Regina

doi:10.1590/1809-4392201601383

Cited by 8 publications

(4 citation statements)

References 4 publications

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“…EDTA remains as anticoagulant of choice for cell count and morphology analysis22 as LH exerts a thrombocyte clumping of the samples 23 24. This effect may be species‐dependent as no cellular morphological differences between K 2 EDTA and LH stored blood samples obtained from fish and parrots were noted 24 25. Both EDTA and LH have an inhibitory effect on bacterial growth26 27 and therefore are not indicated for the storage of samples intended for bacterial culture analysis.…”

Section: Discussionmentioning

confidence: 99%

Concentration of dipotassium ethylenediaminetetraacetic acid but not lithium heparin affects total protein determination in equine synovial fluid

Rihuete¹,

Villarino

Pelisiak

et al. 2020

Veterinary Record

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BackgroundRefractometric determination of total protein (TP) in synovial fluid (SF) is commonly used for diagnosis and monitoring of synovial sepsis in horses. Previous studies have shown that elevated concentrations of certain anticoagulants may overestimate refractometric determination of TP concentration.ObjectivesThe aim of the study was to evaluate the effect of different concentrations of dipotassium EDTA (K2EDTA) and lithium heparin (LH) on TP determination by using a hand-held refractometer in equine synovial fluid.Study designCross-section observational study.MethodsThirty samples of synovial fluid obtained from 22 horses with different synovial conditions were collected. Synovial fluid samples were separated into different aliquots and placed in commercially available collection tubes containing K2EDTA or LH at four different concentrations (1.76, 3.52, 7.04 and 17.6 mg/ml for K2EDTA; 16, 32, 64 and 160 IU/ml for LH) . Refractometric TP determination was performed on untreated and K2EDTA and LH aliquots with a hand-held refractometer and by spectophotometric Biuret method as the gold standard.ResultsRefractometric TP determination was overestimated in SF samples containing 10 times the recommended K2EDTA concentrations. Lower concentrations of K2EDTA and LH concentrations did not affect refractometric TP determinations.Main limitationsLimited number of samples mostly obtained from large synovial structures.ConclusionTo avoid incorrect TP determination, the use of LH containing collection tubes may be an appropriate alternative when the SF volume available is not enough to fill the K2EDTA collection tube.

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Section: Discussionmentioning

confidence: 99%

Concentration of dipotassium ethylenediaminetetraacetic acid but not lithium heparin affects total protein determination in equine synovial fluid

Rihuete¹,

Villarino

Pelisiak

et al. 2020

Veterinary Record

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“…The quantification of plasma protein was done as described by Pessoa et al [ 18 ]: 250 µ l plasma was mixed with 1 mL of Biuret reagent. The mixture was incubated for 30 min and the absorbance was read at 540 nm.…”

Section: Methodsmentioning

confidence: 99%

“…After 5 minutes of incubation, 350 µl of 0.1% naphthyl ethylenediamine was added, additional 5 minutes incubation was observed, and the absorption of the chromophore was read at 546 nm. e quantification of plasma protein was done as described by Pessoa et al [18]: 250 µl plasma was mixed with 1 mL of Biuret reagent. e mixture was incubated for 30 min and the absorbance was read at 540 nm.…”

Section: Biochemical Analysismentioning

confidence: 99%

Antihypertensive Effects of the Methanol Extract and the Ethyl Acetate Fraction from Crinum zeylanicum (Amaryllidaceae) Leaves in L-NAME-Treated Rat

Ndjenda

Nguelefack‐Mbuyo

Atsamo

et al. 2021

Evidence-Based Complementary and Alternative Medicine

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Arterial hypertension (AHT) is a leading cardiovascular disease, with a high negative impact on the quality of life. Crinum zeylanicum (C. zeylanicum) leaves extract is used in the West region of Cameroon to treat AHT and heart problems. This study aimed to investigate the antihypertensive effect of C. zeylanicum extract in Nω-nitro-L‐arginine methyl ester- (L‐NAME-) induced hypertensive rats. The aqueous extract of C. zeylanicum (LAE) was obtained by lyophilizing the juice of triturated fresh leaves. The methanol extract (LME) prepared by maceration of the dried leaves was further partitioned to chloroform (LCF), ethyl acetate (LEAF), and residual (LRF) fractions. The total polyphenol, flavonoid content, and antiradical potentials of these extracts were determined. The curative antihypertensive and renal function protective effects of LME and LEAF were evaluated in vivo on L-NAME-induced hypertensive rats. Hypertension was induced in rats by oral administration of L-NAME (30 mg/kg/day) for 3 consecutive weeks. Thereafter, plant extracts were administered orally at the doses of 30, 60, and 120 mg/kg/day, concomitantly with L-NAME for three other weeks. Body weight, heart rate, and arterial blood pressure were measured at the end of each week throughout the experimental period. At the end of the treatment, 24-hour urine and plasma were collected to assay nitric oxide (NO), creatinine, and protein. The results revealed that LEAF has the higher content of total polyphenol and flavonoid and exhibited the best antiradical potential. Moreover, treatment of hypertensive rats with LME and LEAF significantly ( p < 0.001 ) reduced AHT and heart rate. LME and LEAF significantly increased rat’s body mass, plasmatic NO, and urinary creatinine and reduced urine NO and protein contents as compared to the L-NAME group. LME and its LEAF possess potent antihypertensive effects and further protect the renal function in L-NAME-induced hypertensive rats, thus supporting the use of C. zeylanicum in the management of AHT.

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“…The lipid peroxidation indicator, plasma MDA was measured using thiobarbituric acid reactive substances (TBARS) assay as described previously (Borges et al 2018). Total protein concentration in the serum was measured using protein biuret reaction (Pessoa et al 2017 (Arora et al 2009). Increased in free radical generation inactivates prostacyclin and NO, hence causing half-life of prostacyclin and NO to be decreased.…”

Section: Determination Of Plasma Malondialdehyde (Mda)mentioning

confidence: 99%

Antihypertensive Effect of Piper sarmentosum in L-NAME-Induced Hypertensive Rats

Alwi

Zakaria²,

Karim

et al. 2018

JSM

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Hypertension is one of the risk factors for cardiovascular diseases and has been associated with about 13% of global deaths worldwide. Oxidative stress and reduced nitric oxide (NO) bioavailability contribute to the development of endothelial dysfunction and subsequently hypertension. N ɷ-nitro-L-arginine methyl ester hydrochloride (L-NAME) inhibits NO synthesis; leading to hypertension. Piper sarmentosum (PS) is an herb with antioxidant, antiatherosclerosis and antiinflammation properties. PS also stimulated NO production by endothelial cells. The aim of this study was to determine the effects of aqueous extract of Piper sarmentosum (AEPS) on blood pressure, oxidative stress and the level of nitric oxide in L-NAMEinduced hypertensive rats. Hypertension was induced by oral administration of L-NAME (100 mg/L) in drinking water for four weeks. The rats were concurrently treated with AEPS by oral gavage in serial doses (125, 250 and 500 mg/kg/day). Blood pressure was measured using non-invasive tail-cuff method at baseline and fortnightly thereafter. Serum level of NO and an oxidative stress marker, malondialdehyde (MDA) were measured at baseline and at the end of treatment. The results showed that treatment with three different doses of AEPS successfully reduced systolic blood pressure (p<0.001), diastolic blood pressure (p<0.05) and mean arterial pressure (p<0.05) in L-NAME-induced hypertensive rats. Treatment with AEPS also reduced MDA level (p<0.001) and increased serum NO (p<0.001) in L-NAME-induced hypertensive rats. The findings showed that AEPS decreased blood pressure by protecting against oxidative stress and increasing NO in L-NAME-induced hypertensive rats.

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Refractometry as an alternative to the biuret method for measuring total serum proteins in Podocnemis expansa (Podocnemididae) and Phrynops geoffroanus (Chelidae)

Cited by 8 publications

References 4 publications

Concentration of dipotassium ethylenediaminetetraacetic acid but not lithium heparin affects total protein determination in equine synovial fluid

Concentration of dipotassium ethylenediaminetetraacetic acid but not lithium heparin affects total protein determination in equine synovial fluid

Antihypertensive Effects of the Methanol Extract and the Ethyl Acetate Fraction from Crinum zeylanicum (Amaryllidaceae) Leaves in L-NAME-Treated Rat

Antihypertensive Effect of Piper sarmentosum in L-NAME-Induced Hypertensive Rats

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