Labmax 240

The biochemical analyses were performed twice: during the selection of the participants and after the end of the nutritional intervention. In both analyses, the levels of total cholesterol (TC) and its fractions (low-density lipoprotein (LDL) and high-density lipoprotein (HDL)), triglycerides, malondialdehyde (MDA), total antioxidant capacity (TAC), homocysteine, and vitamin B12 were measured. The blood samples were collected after a 12-h fast at home, using sterile vacuum tubes with and without anticoagulant, according to the guidelines for the use of sharp materials.
Lipid profile concentrations were determined using the turbidimetry method using a Labmax 240 premium-Labtest automated biochemical analyzer. For analysis of antioxidant activity through the plasma MDA and serum TAC, we used a previously described protocol [18 (link)]. Homocysteine levels were quantified using the high-performance liquid chromatography (HPLC) method [19 (link)]. The serum concentrations of folic acid and vitamin B12 were measured by chemiluminescence and an electrochemiluminescence immunoassay, respectively.

Biochemical measurements of blood and urine samples were collected after a 12 h fast and were analyzed at the Vizela Laboratory of the Municipal Health Foundation and at the Antônio Pedro University Hospital (HUAP) of UFF. Biochemical evaluations performed using the Wiener^® Selectra equipment were: glucose (Gli) by hexoquinase method; insulin by chemiluminescence; total cholesterol (TC) and triglycerides (TG) by enzymatic kinetics; and HDL-cholesterol (HDL-c) and LDL-c by enzymatic colorimetric assay. HbA1c was measured by immunoturbidimetry using a Labmax 240 equipment from Labtest. Sample collection and biochemical analyses took place on the same day. Blood pressure was measured three times with a Pro Check^® digital manometer and a standardized oscillometric blood pressure device (OmronHealthcare Incorporated, Lake Forest, USA), allowing for calculation of the mean value between the second and third measurements [22 (link)]. When the difference was higher than 5 mm Hg between measurements, a new measurement was carried out.

Serum levels of alanine aminotransferase (ALT; IU/L), aspartate aminotransferase (AST; IU/L), alkaline phosphatase (ALP; IU/L), conjugated and total bilirubin (mg/dL) were measured with an automated spectrophotometric Labmax 240 analyzer (Labtest Diagnostica, Lagoa Santa, Brazil) after appropriate dilution of samples.

Blood, collected from inferior vena cava 6, 12 and 24 hours after injection, was centrifuged (10 minutes, 1500 × g), and serum was stored at -20°C. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured with an automated spectrophotometric analyzer (Labmax 240, Labtest Diagnostica, Lagoa Santa, MG, Brazil). Values were expressed in IU/L.

Alanine aminotransferase (ALT) (IU/L), aspartate aminotransferase (AST) (IU/L), alkaline phosphatase (ALP) (IU/L) and conjugated bilirubin (mg/dL) values were measured with an automated spectrophotometric Labmax 240 analyzer (Labtest Diagnostica, Brazil) or an automated bench-top dry chemistry analyzer (IDEXX Laboratories Ltd, UK) after appropriate dilution of collected serum samples.

Approximately 20 ml of blood were collected during admission at the hospital, and aliquots of both plasma and serum was stored at -80 °C for later measurements. All analyses were performed in the automatic LABMAX 240® equipment from Labtest, using commercial standards for Albumin (LABTEST), C-Reactive Protein (LABTEST), and hemoglobin (LABTEST).

The clinical evaluation included inspection for jaundice, lymphadenopathy and hyperthermia. The shelter’s employees were instructed to report any gastrointestinal, urogenital, cardio-respiratory, nervous or behavioral disorders.
Serum biochemistry analysis was performed in a Labmax 240 device (Labtest Diagnostica, Minas Gerais, Brazil) using an Enzymatic Kinetic Method kit (Rx Series, Randox, Crumlin, UK), following the manufacturer’s specifications. The analysis included alkaline phosphatase (ALP) and alanine aminotransferase activity (ALT). Total bilirubin (TB), total protein (TP), blood urea nitrogen (BUN) and creatinine (CR) serum concentrations were also determined. Hematological analysis was performed in an ABX Micros ABC Vet (Horiba Medical, Kyoto, Japan) within four hours of sampling and included white blood cell count (WBC), red blood cell count (RBC) and platelet count (PLT). The differential WBC count was performed by optical microscopy of Rosenfeld-stained blood smears when necessary (Modified May-Grünwald). The reference intervals adopted for this study are presented in S1 Appendix.