cometadata/datacite-titles-descriptions-related-identifiers huggingface dataset:
DOI: 10.7910/dvn/zlf8og
title: A16_32439.jpg

description: Link to OCHRE database: &lt;a href = 'http://pi.lib.uchicago.edu/1001/org/ochre/0bdbaffb-dd08-4af7-a677-ec82b90a8405'&gt;http://pi.lib.uchicago.edu/1001/org/ochre/0bdbaffb-dd08-4af7-a677-ec82b90a8405
DOI: 10.7910/dvn/ajsdqv
title: A16_32440.jpg

description: Link to OCHRE database: &lt;a href = 'http://pi.lib.uchicago.edu/1001/org/ochre/6467acac-d2b1-4871-ab9f-593573827abf'&gt;http://pi.lib.uchicago.edu/1001/org/ochre/6467acac-d2b1-4871-ab9f-593573827abf
DOI: 10.57715/unr/5xifxn/acvjky
title: 2022-02-02_Growth curves 1_ Starved cells plus Cu 50 a 250.tab

description: Growth curves_ General Method 
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were routinely maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L Liver Infusion Broth BD DIFCO™ # 226920, 5 g/L GranuCult® prime Tryptose Broth Millipore #1.03866, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS (Internegocios S.A. - Buenos Aires, Argentina) and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C.
Cell growth was determined by cell counting in an automatized hemocytometer adapted to count epimastigotes (WL 19 Counter AA, Weiner Lab) and in Neubauer counting chamber.


Growth curves - set 1
Epimastigotes were cultured in LIT-10 % FBS-0 μM hemin medium for at least 10 days before the performance of the experiment shown in this set of growth curves. For the experiment, the cells were diluted at a concentration of 5 x 106 parasites/mL and challenged to grow in LIT-10 % FBS-0 μM hemin supplemented with 50-250 μM Cu. Control was performed by adding 5 µM hemin to the same medium (without copper). The number of cells was monitored for 7 days.
Copper (Cu) was added as Cu2+ from CuSO4 salt.

Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/kezgbx
title: 2022-02-11_Growth curves 2_Copper trearment.tab

description: Growth curves_ General Method 
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were routinely maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L Liver Infusion Broth BD DIFCO™ # 226920, 5 g/L GranuCult® prime Tryptose Broth Millipore #1.03866, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS (Internegocios S.A. - Buenos Aires, Argentina) and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C.
Cell growth was determined by cell counting in an automatized hemocytometer adapted to count epimastigotes (WL 19 Counter AA, Weiner Lab) and in Neubauer counting chamber.


Growth curves – set 2
Routinely maintained cells were diluted at a concentration of 5 x 106 parasites/mL and challenged to grow in LIT-10 % FBS-5 μM hemin medium supplemented with 50-500 μM Cu. The number of cells was monitored for 6 days.
Copper (Cu) was added as Cu2+ from CuSO4 salt.

Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.7910/dvn/379pux
title: A16_32443.jpg

description: Link to OCHRE database: &lt;a href = 'http://pi.lib.uchicago.edu/1001/org/ochre/b2066885-a169-452a-9252-89da84d2f33b'&gt;http://pi.lib.uchicago.edu/1001/org/ochre/b2066885-a169-452a-9252-89da84d2f33b
DOI: 10.57715/unr/5xifxn/jcstyl
title: 2021-12-30al 1-3_Viability test_Cu and NeoCup.tab

description: 2021-12-30al 1-3_Viability test_Cu and NeoCup
Vero cells were plated on 96 multi-well plate in DMEM-2 % FBS (4 x 103 cells per well) and incubated for 24 hours. Then, the cells were washed with PBS and incubated with 200 μL of DMEM-2 % FBS supplemented with 50-500 μM Cu and 0.01-5 μM NeoCup. No addition was made in the 100 % viability control. After 72 h, the viability of cells was measured by MTT reduction colorimetric method as described (https://doi.org/10.1515/cclm.1995.33.11.813). Briefly, the cells were incubated in 180 μL DMEM-2 % FBS plus 20 μL 5 mg/mL MTT for 1 h, washed with PBS and mixed with 100 μL DMSO. Absorbance at 540 nm was measured to determine formazan production.
Reagents
Dulbecco’s Modified Eagle Medium (DMEM) was obtained from Life Technologies. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min.
Copper (Cu) was added as Cu2+ from CuSO4 salt. Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate.

 Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/kaf5ku
title: 2022-01-17_Effect of Cu and chelators in the infection_3rd.tab

description: Infection assay:
Vero cells (ATCC CCL-81) were seeded on 12 mm round coverslips in 24-well culture plates, at a density of 1.58 x 103 cells/cm2, and incubated in DMEM- 2 % FBS. After 24 h, they were infected with cell-derived trypomastigotes (MOI of 10) for 16 h in DMEM-2 % FBS supplemented with 50-150 μM Cu, 50-500 μM BCS, or 0.005-0.02 μM NeoCup. Then, non-internalized trypomastigotes were washed twice with PBS and the medium was renewed. 72 h post infection, the monolayers of Vero cells were washed with PBS, fixed with methanol for 15 min, washed with stabilized water, and stained with Giemsa’s reagent for 30 min. Finally, they were washed with stabilized water, air dried, and mounted with Canada balsam. 200-300 cells per slide were analyzed by light microscopy to determine the number de amastigotes per cell. 0 indicates a cell without amastigotes.
Reagents: 
Dulbecco’s Modified Eagle Medium (DMEM) was obtained from Thermo Fisher Scientific (#12100046). Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min.
Copper (Cu) was added as Cu2+ from CuSO4 salt. Bathocuproine disulfonate (BCS, an extracellular copper chelator). Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate (Supelco # 72090).
 Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/kmkhbu
title: 2022-08-25_Effect of Cu and chelators on metaciclogenesis.tab

description: Quantification of metacyclogenesis:
Epimastigotes were maintained in mid-log phase by dilutions every 2-3 days in LIT-10 % FBS 5 μM hemin medium, at least for 10 days. Then, the cells were washed with PBS, diluted at a concentration of 5 x 106/mL and incubated in LIT-10 % FBS 5 μM hemin pH 6 medium for 7 days at 28 °C. The medium was supplemented with Cu, BCS or NeoCup (concentration in μM). The cells were collected, washed with PBS, fixed with 3.7 % (w/v) formaldehyde in PBS, and washed with PBS. Then, the number of epimastigotes and metacyclic trypomastigotes were counted in Neubauer chamber. 
Reagents:
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L liver infusion, 5 g/L bacto-tryptose, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min. Hemin stock solution (1 mM) was prepared in 50 % (v/v) EtOH, 0.01 N NaOH, fractionated and stored at −80 °C.
Copper (Cu) was added as Cu2+ from CuSO4 salt. Bathocuproine disulfonate (BCS, an extracellular copper chelator). Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate.
 Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/nhcciz
title: 2022-11-08_ROS test_in triplicates_3rd.tab

description: Superoxide anion production in epimastigotes
Superoxide anion production was quantified as it was previously described (https://doi.org/10.1016/j.cryobiol.2013.05.005) with some modifications. Epimastigotes were maintained for 10 days in mid-log growth phase by periodic dilutions in LIT-10 % FBS medium without hemin. Then, the cells were diluted at a concentration of 5 x 106 parasites/mL and challenged to grow in LIT-10 % FBS medium without hemin plus 100 µM Cu (or no addition for controls). After 5 days, 50 x 106 cells per sample were collected, washed with PBS, resuspended in 270 µL PBS plus 30 µL of 10 mg/mL NBT (nitro-blue tetrazolium chloride), and incubated 30 min at 28 °C. After the incubation, the cells were washed with PBS and lysed with 161.6 µL DMSO. The mix was vortexed and then 138.4 µL 2 M NaOH were added. Absorbances of the samples were measured in microplate reader (Multimodal Microplate Reader, model Synergy HTX version S1LFA, BioTek Instruments) at 540 nm (the concentration of superoxide anion can be calculated using an extinction coefficient of 7.2 cm2 µmol−1).
Reagents
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L liver infusion broth, 5 g/L bacto-tryptose, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C. Liver Infusion Broth BD DIFCO™ # 226920. GranuCult® prime Tryptose Broth Millipore #1.03866. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min. Hemin (Frontier Scientific #H651-9) stock solution (1 mM) was prepared in 50 % (v/v) EtOH, 0.01 N NaOH, fractionated and stored at −80 °C.
Copper (Cu) was added as Cu2+ from CuSO4 salt. Bathocuproine disulfonate (BCS, an extracellular copper chelator). Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate (Supelco # 72090).

 Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/o1l3hx
title: 2022-09-09_Growth curves 3_BCS and NeoCup treatment.tab

description: Growth curves_ General Method 
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were routinely maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L Liver Infusion Broth BD DIFCO™ # 226920, 5 g/L GranuCult® prime Tryptose Broth Millipore #1.03866, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS (Internegocios S.A. - Buenos Aires, Argentina) and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C.
Cell growth was determined by cell counting in an automatized hemocytometer adapted to count epimastigotes (WL 19 Counter AA, Weiner Lab) and in Neubauer counting chamber.


Growth curves – set 3
Routinely maintained cells were diluted at a concentration of 5 x 106 parasites/mL and challenged to grow in LIT-10 % FBS-5 μM hemin medium supplemented with 50-500 μM BCS or 0.01-100 μM NeoCup. The number of cells was monitored for 6 days.
Bathocuproine disulfonate (BCS, an extracellular copper chelator). Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate (Supelco # 72090).


Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/vmmmbi
title: 2021-12-23_27_Viability test_BCS.tab

description: 2021-12-23_27_Viability test_BCS
Vero cells were plated on 96 multi-well plate in DMEM-2 % FBS (4 x 103 cells per well) and incubated for 24 hours. Then, the cells were washed with PBS and incubated with 200 μL of DMEM-2 % FBS supplemented with 50-500 μM BCS. No addition was made in the 100 % viability control. After 72 h, the viability of cells was measured by MTT reduction colorimetric method as described (https://doi.org/10.1515/cclm.1995.33.11.813). Briefly, the cells were incubated in 180 μL DMEM-2 % FBS plus 20 μL 5 mg/mL MTT for 1 h, washed with PBS and mixed with 100 μL DMSO. Absorbance at 540 nm was measured to determine formazan production.
Reagents
Dulbecco’s Modified Eagle Medium (DMEM) was obtained from Life Technologies. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min.
Bathocuproine disulfonate (BCS, an extracellular copper chelator). 


 Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/fknkoz
title: 2022-09-30_Oxygen consumption with Cu_BCS_NeoCup_day3.tab

description: 2022-09-30_Oxygen consumption with Cu_BCS_NeoCup_day3
Oxygen consumption was measured in epimastigotes grown in LIT-10 % FBS-5 μM hemin medium plus copper (Cu), BCS, NeoCup or no addition (control), for 3 days. Concentration in μM.
The oxygen consumption rates from epimastigotes were quantified from the linear response, as previously described*, using a Clark electrode connected to a 5300 Biological Oxygen Monitor (Yellow Springs Instrument Co.). Epimastigotes were collected, washed with PBS, suspended in TSB-EGTA (125 mM sucrose, 65 mM KCl, 1 mM MgCl2, 2.5 mM KH2PO4, 10 mM Tris-HCl pH 7.5, and 333 μM EGTA) at a ratio of 5 × 105 cells/μL and kept in ice. Before measurements, 5 mM Succinate and 50 μM digitonin were added to each sample. For oxygen consumption, 50-150 μL of permeabilized parasites were added to 1.9 mL of TSB-EGTA plus 5 mM succinate at 28 °C.
* Vercesi AE, Bernardes CF, Hoffmann ME, Gadelha FR, Docampo R. Digitonin permeabilization does not affect mitochondrial function and allows the determination of the mitochondrial membrane potential of Trypanosoma cruzi in situ. J Biol Chem. 1991 Aug 5;266(22):14431-4. PMID: 1860850.


Reagents
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L liver infusion, 5 g/L bacto-tryptose, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min. Hemin stock solution (1 mM) was prepared in 50 % (v/v) EtOH, 0.01 N NaOH, fractionated and stored at −80 °C.
Copper (Cu) was added as Cu2+ from CuSO4 salt. Bathocuproine disulfonate (BCS, an extracellular copper chelator). Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate.
 Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/mofro0
title: 2024-06-11_Viability test on epimastigotes.tab

description: T. cruzi epimastigotes cells were diluted at a concentration of 5 x 106 parasites/mL and challenged to grow in LIT-10 % FBS-5 μM hemin medium supplemented with 0.02, 0.5, and 1 μM NeoCup. After 3 or 7 days, cells were stained with Erythrosin B Stain 0.5% and the percentage of cell viability was determined. Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate (Supelco # 72090). Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/podb7r
title: 2022-10-17_Oxygen consumption with BCS_day 5.tab

description: 2022-10-17_Oxygen consumption with BCS_day 5
Oxygen consumption was measured in epimastigotes grown in LIT-10 % FBS-5 μM hemin medium plus BCS or no addition (control), for 5 days. Concentration in μM.
The oxygen consumption rates from epimastigotes were quantified from the linear response, as previously described*, using a Clark electrode connected to a 5300 Biological Oxygen Monitor (Yellow Springs Instrument Co.). Epimastigotes were collected, washed with PBS, suspended in TSB-EGTA (125 mM sucrose, 65 mM KCl, 1 mM MgCl2, 2.5 mM KH2PO4, 10 mM Tris-HCl pH 7.5, and 333 μM EGTA) at a ratio of 5 × 105 cells/μL and kept in ice. Before measurements, 5 mM Succinate and 50 μM digitonin were added to each sample. For oxygen consumption, 50-150 μL of permeabilized parasites were added to 2 mL of TSB-EGTA plus 5 mM succinate at 28 °C.
* Vercesi AE, Bernardes CF, Hoffmann ME, Gadelha FR, Docampo R. Digitonin permeabilization does not affect mitochondrial function and allows the determination of the mitochondrial membrane potential of Trypanosoma cruzi in situ. J Biol Chem. 1991 Aug 5;266(22):14431-4. PMID: 1860850.

Reagents
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L liver infusion, 5 g/L bacto-tryptose, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min. Hemin stock solution (1 mM) was prepared in 50 % (v/v) EtOH, 0.01 N NaOH, fractionated and stored at −80 °C.
Bathocuproine disulfonate (BCS, an extracellular copper chelator).

 Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/spzslb
title: 2023-06_ICP.tab

description: ICP-OES
Cu, Fe, Ca, K, Mg, Mn, Na, P, S, and Zn contents were determined by ICP-OES as described in (https://doi.org/10.7554/eLife.64690). Briefly, epimastigotes were maintained for 6 days in LIT-10 % FBS-5 μM hemin medium plus different concentrations of copper or copper chelators, or in LIT-10 % FBS without hemin medium plus different concentrations of hemin or iron. For heme starved cultures, the parasites were maintained the 10 previous days at mid-log phase by periodic dilutions in LIT-10 % FBS without hemin medium before the treatments. 120 x 106 epimastigotes were collected per measurement and lyophilized. The samples were digested by boiling for 1 h in 40 % nitric acid, then diluted in ultra-pure metal-free water and analyzed by ICP-OES (Perkin Elmer, Optima 7300DV) versus acid-washed blanks. Concentrations were determined from a standard curve constructed with serial dilutions of two commercially available mixed metal standards (Optima). Blanks of nitric acid with and without ‘metal-spikes’ were analyzed to ensure reproducibility.

Reagents
The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L liver infusion, 5 g/L bacto-tryptose, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min. Hemin stock solution (1 mM) was prepared in 50 % (v/v) EtOH, 0.01 N NaOH, fractionated and stored at −80 °C.
Copper (Cu) was added as Cu2+ from CuSO4 salt. Bathocuproine disulfonate (BCS, an extracellular copper chelator). Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate.

 Authors: Merli, Marcelo Luciano; Xinyu Zhu, Paul A. Cobine; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/zcopw1
title: 2024-04 to 06_ICP samples_Epis with Neo.tab

description: ICP-OES Cu, Fe, Ca, K, Mg, Mn, Na, P, S, and Zn contents were determined by ICP-OES as described in (https://doi.org/10.7554/eLife.64690). Briefly, epimastigotes were maintained for 6 days in LIT-10 % FBS-5 μM hemin medium plus different concentrations of NeoCup. Around 100-120 x 106 epimastigotes were collected per measurement and lyophilized. The samples were digested by boiling for 1 h in 40 % nitric acid, then diluted in ultra-pure metal-free water and analyzed by ICP-OES (Perkin Elmer, Optima 7300DV) versus acid-washed blanks. Concentrations were determined from a standard curve constructed with serial dilutions of two commercially available mixed metal standards (Optima). Blanks of nitric acid with and without ‘metal-spikes’ were analyzed to ensure reproducibility. Reagents The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L liver infusion, 5 g/L bacto-tryptose, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min. Hemin stock solution (1 mM) was prepared in 50 % (v/v) EtOH, 0.01 N NaOH, fractionated and stored at −80 °C. Neocuproine solution (NeoCup, an intracellular copper chelator) was prepared from Neocuproine hydrochloride monohydrate. Authors: Merli, Marcelo Luciano; Xinyu Zhu, Paul A. Cobine; Cricco, Julia Alejandra.
DOI: 10.57715/unr/5xifxn/e3glr3
title: 2022-11-08_Intracelular heme.7z

description: The file “2022-11-08_Intracelular heme/2022-11-08_Samples.xlsx” shows the sample list for Heme content determination. The spectrums are recorded in separate files named as "PVC WAVE-SCAN 2022-11-08 ..." (22 files). Method: Intracellular heme content determined by pyridine method in epimastigotes grown in LIT-10 % FBS-5 μM hemin medium plus 100 μM Cu for 4 days, plus 500 μM Cu for 6 days or without addition (control). The heme content was measured by the alkaline pyridine method described in (10.1016/0003-2697(87)90643-9). We made modifications to perform the assay with epimastigotes (https://doi.org/10.1371/journal.pntd.0004359). Briefly, 50-200 x 106 epimastigotes were collected, washed twice with PBS, and suspended in 500 μL PBS. The sample was combined with a volume of 500 μL containing 0.2 M NaOH and 40 % (v/v) pyridine. Reduced spectra were recorded by addition of sodium dithionite. Heme concentration should be calculated from the differential -reduced minus oxidized- spectrum. Molar extinction coefficient at 557 nm: 23.98 mM−1 cm−1. Reagents: The experiments were done using T. cruzi Dm28c strain. T. cruzi epimastigotes were maintained in mid-log phase by periodic dilutions in Liver Infusion Tryptose (LIT) medium (5 g/L liver infusion, 5 g/L bacto-tryptose, 68 mM NaCl, 5.3 mM KCl, 22 mM Na2HPO4, and 0.8 % (w/v) glucose, pH 7.4) supplemented with 10 % FBS and 5 μM hemin (LIT-10 % FBS-5 μM hemin medium), at 28 °C. Fetal Bovine Serum (FBS) was obtained from Internegocios S.A. (Buenos Aires, Argentina) and heat-inactivated at 56 °C for 30 min. Hemin stock solution (1 mM) was prepared in 50 % (v/v) EtOH, 0.01 M NaOH, fractionated and stored at −80 °C. Copper (Cu) was added as Cu2+ from CuSO4 salt. Authors: Merli, Marcelo Luciano; Cricco, Julia Alejandra.
DOI: 10.15468/dl.yj92p6
title: Occurrence Download

description: A dataset containing 6597 species occurrences available in GBIF matching the query:
{
 "and" : [
 "HasGeospatialIssue is false",
 "HasCoordinate is true",
 "TaxonKey is one of (Plethodon teyahalee Hairston, 1950)"
 ]
}

 The dataset includes 6597 records from 11 constituent datasets; see https://api.gbif.org/v1/occurrence/download/0003679-250415084134356/datasets/export for details.
 
Data from some individual datasets included in this download may be licensed under less restrictive terms.
DOI: 10.15468/dl.wmdgss
title: Occurrence Download

description: A dataset containing 608 species occurrences available in GBIF matching the query:
{
 "and" : [
 "BasisOfRecord is Human Observation",
 "HasCoordinate is true",
 "HasGeospatialIssue is false",
 "TaxonKey is Osmia parietina",
 "Year 2000-2025"
 ]
}

 The dataset includes 608 records from 39 constituent datasets; see https://api.gbif.org/v1/occurrence/download/0007032-250827131500795/datasets/export for details.
 
Data from some individual datasets included in this download may be licensed under less restrictive terms.