Edta tube

This study involved typing 79 P. vivax-exposed individuals from the aforementioned endemic areas for the HLA-DRβ1 allele; PB samples from 50 individuals living in Bogotá (a non-endemic malaria area) who have never been exposed to malaria were also taken for typing to form the control group. PB samples were taken from P. vivax-exposed individuals and the control group and placed in EDTA tubes (BD Vacutainer Oakville, ON, CAN). A Wizard Genomic DNA Purification Kit (Promega Corporation, Madison, WI, USA) was used for extracting genomic DNA (gDNA) from 300 μL whole blood, according to the manufacturer’s instructions. The gDNA was verified by 1% agarose gel with SYBR Safe DNA Gel Stain (Thermo Fisher Scientific, CA, USA) and sent to Histogenetics (Ossining, NY, USA) for high-resolution HLA sequence-based typing (SBT) of the β1 locus.

Blood samples were taken from the auricular vein at week 14 and 28. After 7 hours fasting, samples were collected in EDTA tubes (BD Vacutainer, Plymouth, UK), stored on ice and centrifuged (1500 g, 15 min, 4^°C). Then, plasma was obtained and stored in a -80^°C refrigerator. Plasma triglycerides, total cholesterol, HDL, LDL, transaminases, gamma-glutamyl transpeptidase (GGT), bile acid, bilirubin, creatinine, urea, total protein, albumin, glucose and creatine phosphokinase (CPK) were determined using standard enzymatic procedures by an external laboratory (Immunovet, Barcelona, Spain).

Ten mL of peripheral blood was collected in EDTA tubes (BD Vacutainer® Franklin Lakes, NJ) from three mCRPC patients before initiation of CRXL301 chemotherapy (baseline; cycle 1 day 1; C1D1). Additionally, 10 mL of peripheral blood was also collected at the following time points: C1D1 plus four hours, C1D2, C1D8, C2D1, C2D1 plus four hours, and C2D8.
Blood samples were shipped at ambient temperature on same day of collection and were processed within 24 h from blood draw. Samples were processed as previously described^{[22 (link)]}.
Briefly, CTCs were enriched by negative depletion of CD45⁺ cells (peripheral mononuclear cells, PBMCs) by using the RosetteSep^TM Human CD45 Depletion Cocktail (Stemcell Technologies, Cambridge, MA); the enriched CTCs were subsequently cytospun onto coverslips and processed for immunofluorescence staining.

Venous blood of healthy women was collected into 10 mL EDTA tubes (BD Vacutainer; Franklin Lakes, NJ, USA). Polymorphonuclear cell (PMN) isolation was performed by positive selection using CD15 MicroBeads (Miltenyi Biotec; Auburn, CA, USA) and a whole blood column kit following the manufacturer’s instructions (Miltenyi Biotec). This isolation method was effective with 92.84% viability (Figure 1a and Figure S1a) and >94% neutrophil enrichment (as CD45⁺ CD15⁺ CD66b⁺ cells), determined by flow cytometry. Purified neutrophils were resuspended in HBSS culture medium (Hanks’ Balanced Salt Solution, Gibco; Waltham, MA, USA) for imaging analysis or in X-VIVO 15 media for cell culture (Lonza; Bend, OR, USA) and stimulated with 25 mM of sodium acetate, sodium butyrate or sodium propionate for 1 or 3 h as indicated for further analysis.

Whole blood was collected from both patients in EDTA tubes (BD Vacutainer, San Diego, CA). Samples from patient 1 were obtained at three different points (T0 = pre-transplantation, T1 = 1 day post-transplantation, and T7 = 7 days post-transplantation). PBMCs were isolated by density gradient centrifugation on Ficoll (Cedarlane Laboratories Limited, Hornby, ON, Canada) and viable cells were counted with the automated cell counter ADAM-MC (Digital Bio, NanoEnTek Inc., Seoul, KR, Korea). PBMCs were resuspended at the concentration of 1 × 10⁶/mL in RPMI 1640 medium (Euroclone, Milan, Italy) supplemented with 10% fetal bovine serum, 1% of L-glutamine (LG), and 2% pen-streptomycin. Subsequently, PBMCs were stimulated with 500 ng/mL nucleocapsid- (N) and spike- (S) specific SARS-CoV-2 antigens (Novatein Biosciences, Cambridge, MA, USA). For gene expression and cytokine analyses, cells were harvested 10 h after stimulation and cytokine content was quantified on cell culture supernatants.