eamag/protocols-with-steps huggingface dataset:
TITLE: Importing live fungi to the US (UF Forest Entomology Lab)
AUTHORS: Demian F Gomez, You Li, Jiri Hulcr

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes how to hand-carry live fungi through airport customs into the United States for the UF Forest Entomology Lab.</div><div class = "text-block"><span>This protocol is part of the Bark Beetle Mycobiome (BBM) Research Coordination Network. For more information on the BBM international network: Hulcr J, Barnes I, De Beer ZW, Duong TA, Gazis R, Johnson AJ, Jusino MA, Kasson MT, Li Y, Lynch S, Mayers C, Musvuugwa T, Roets F, Seltmann KC, Six D, Vanderpool D, & Villari C. 2020. Bark beetle mycobiome: collaboratively defined research priorities on a widespread insect-fungus symbiosis. Symbiosis 81: 101–113 </span><a href="https://doi.org/10.1007/s13199-020-00686-9" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">https://doi.org/10.1007/s13199-020-00686-9</span></a><span>.</span></div></div>

[STEPS]
TITLE: Statistical Considerations and Analysis Plan (Part 9 of Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-onset Type 1 Diabetes Mellitus)
AUTHORS: Stephen.Gitelman , Jeffrey A. Bluestone

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This is Part 9 of "Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-Onset Type 1 Diabetes Mellitus".</div><div class = "text-block">This clinical study is supported by JDRF. The aim of the collection is to determine whether imatinib will slow the progression of the autoimmune destruction of ß cells and lead to the preservation of C-peptide secretion in T1DM and to assess Diabetes-related objectives and safety of Imatinib in new-onset type 1 diabetes mellitus".</div></div>

[STEPS]
TITLE: Nuclei counting after Trypan Blue staining
AUTHORS: Carmen Abaurre, Ka Wai Lee, Ernest Arenas

[DESCRIPTION]
Nuclei counting after Trypan Blue staining

[GUIDELINES]
Be wary of nuclei integrity when counting for snRNA-seq. Some minor blebbing of nuclei can still be considered acceptable, but broken up nuclei are considered debris and should not be counted, as they have lost single-cell characteristics. 
Trypan Blue also stains debris, so for counting it is important to have a clean nuclei suspension.

[STEPS]
1. Prepare nuclei suspension from samples. 
2. In order to increase accuracy, prepare duplicates or triplicates per sample.
3. Add 9 μl of 0.4% Trypan Blue to a 0.2 ml tube
4. Add 1 µl of nuclei suspension to the Trypan Blue solution. Do not dispense to the walls of the tube.
5. Mix slowly by pipetting up and down 10 times. If the nuclei preparation is prone to clumping or losing integrity (breakage and release of chromatin), use a 20 µL pipette and further reduce mixing speed. 
6. Fill the counting chamber. Observe at 20X
7. Calculate the average concentration obtained across duplicates/triplicates of the sample
TITLE: Harvesting OP50 for C. elegans liquid culture
AUTHORS: Vidur Sabharwal

[DESCRIPTION]
<div class = "text-blocks"></div>

[STEPS]
?. [Harvesting OP50]
After 8 hr incubation, use 2 ml of E. coli OP50 LB culture to inoculate each of 3 x 250 ml LB. Place flasks in shaking incubator (180 rpm) O/N.
37 °C
?. [Harvesting OP50]
Weigh 3 x 50 ml centrifuge tubes and write weight of the empty tube on the tube.
?. [Harvesting OP50]
Day 2. Using a serological pipette, aliquot 40 ml of O/N E. coli OP50 culture into the pre weighed centrifuge tubes. Centrifuge in swing bucket centrifuge at 3,220 x g, 10°C, 10 min.
10 °C
?. [Harvesting OP50]
Carefully decant the supernatant, repeat step 4 till all the culture is used up. Keep the tube inverted with lid on and leave to stand for several minutes. Using a pipette remove any excess supernatant that may have collected in the lid. Weigh the tube and calculate the weight of the pellet.
?. [Harvesting OP50]
Calculate the volume needed to provide a suspension of 30 g/L and mark this volume on the tube. Date, label and place tubes at -20 °C for use within 1-3 months or at - 80 °C if storing for longer than 3 months.
?. [Harvesting OP50]
Prepare bacterial suspension for growing nematodes; allow bacterial pellet to thaw out and add required volume of S complete medium to each tube, vortex gently to resuspend pellet, pool contents of different tubes to obtain the volume required. Work under sterile conditions.
TITLE: Determination of Clostridium sp. contamination level
AUTHORS: Eva Petrova, Roey Angel

[DESCRIPTION]
adopted from Dr Tomasz Grenda (2018)

[STEPS]
1. Prepare 10g of sample and resuspend it in 90ml of saline solution (0.85% NaCl). 
10 g 
90 mL 

2. Subject suspension to decimal dilutions in saline solution (see Guidelines), prepare at least 5 levels. 
3. After sample preparation, dispense 10ml of TPGY broth into tubes (number of tubes depends on the number of samples multiplied by prepared dilutions, each tube corresponds to examined level). Transfer 1ml of each dilution to tubes with dispensed and preregenerated TPGY broth. 

10 mL 1 mL 


4. Subject the samples to pasteurization process at 70°C for 15 min. 
70 °C 
15 min 

5. Incubate inoculated tubes at 30°C or 37°C for 48h, under anaerobic conditions.
37 °C 
2880 min 

6.  After incubation check the turbidity of TPGY and the ability to gas production. 
7. Spread 10µl of liquid culture onto surfaces of FAA and Willis–Hobbs agar media. Incubate inoculated plates at 37°C for 48h, under anaerobic conditions. 
10 µL 
37 °C 
2880 min 


8. After incubation evaluate characteristic phenotypic features of obtained cultures and determine the contamination level with clostridia adequately to dilution at which their growth was noticed.
TITLE: Kasson Lab DNA Extraction
AUTHORS: Angie Macias, Matthew T Kasson, Brian Lovett

[DESCRIPTION]
This is a routine protocol for extracting DNA from various fungi. This extraction method is suitable for follow-up molecular work such as PCR amplification.

[STEPS]
SECTION: Before you begin
1. Turn on hot water bath, set to 65 °C.

SECTION: Before you begin
2. Pull two Eppendorf 1.5 mL centrifuge tubes per sample.

SECTION: Before you begin
2.1. Label both sets of tubes with (short) sample names.
SECTION: Before you begin
2.2. Label one tube set for each sample with an "I" for  .
  

SECTION: Before you begin
3. Add 600 µL of   (or () to tubes without "I".

SECTION: Before you begin
4. Add 600 µL of   to tubes labeled with "I".

SECTION: Before you begin
5. Place tube with   into 65 °C water bath.

SECTION: Extraction Protocol
6. Sterilize some metal scrapers with flame and 95 % (v/v)   .

SECTION: Extraction Protocol
7. Add 1/2 pea-sized amount of fungal tissue (young hyphae) to each tube containing  .

SECTION: Extraction Protocol
7.1. Flame-sterilize and cool scrapers between samples.
SECTION: Extraction Protocol
8. Macerate each sample with a new, sterile micropestle until tissue is homogenous.
SECTION: Extraction Protocol
9. Add tubes to a floating rack to allow samples to incubate directly in 65 °C water bath for 30 min.

SECTION: Extraction Protocol
10. Remove samples and vortex for 3 s before returning to 65 °C water bath for 30 min.

SECTION: Extraction Protocol
11. Remove samples and allow them to cool on the bench for 5 min.

SECTION: Extraction Protocol
12. Add 200 µL of   to each tube and vortex for 10 seconds.

SECTION: Extraction Protocol
13. Centrifuge samples for 3 min at 14.000 rpm.

 

SECTION: Extraction Protocol
14. Using a P1000 micropipette, transfer supernatant to each tube containing   and gently mix by inversion.
 


SECTION: Extraction Protocol
15. Centrifuge for 1 min at 14.000 rpm.

SECTION: Extraction Protocol
16. Carefully pour off the supernatant into waste container.
 

SECTION: Extraction Protocol
17. Add 600 µL of 70 % (v/v)   to each tube and mix gently by inversion.

SECTION: Extraction Protocol
18. Centrifuge for 1 min at 14.000 rpm.

SECTION: Extraction Protocol
19. Repeat Step 16.
SECTION: Extraction Protocol
20. Open and invert tubes onto a clean paper towel.
 

SECTION: Extraction Protocol
21. Add 100 µL of warmed   to each tube.

SECTION: Extraction Protocol
22. Store fully-labeled tubes in a box (not a tube rack) in the -20 °C freezer.

SECTION: Extraction Protocol
10.1. Place a sufficient aliquot of   in water bath to warm for Step 21.
TITLE: Purification of influenza A virus RNA from cell supernatant to check sequences
AUTHORS: Marta.Gaglia, lea.gaucherand

[DESCRIPTION]
We use this protocol to isolate RNA from influenza A virus-containing cell supernatant and monitor maintenance of sequence insertions or mutations

[STEPS]
1. Influenza A virus extraction from cell supernatant using QIAamp Viral RNA Mini Kit
Follow the manufacturer's protocol using a starting cell supernatant volume of 140 µl.
Elute in 60 µl buffer AVE.
2. DNase treatment using Qiagen RNase-free DNase set:
Make mastermix of DNase and buffer. For each sample:
10 µl buffer RDD
2.5 µl DNase I
Add to each RNA tube, 27.5 µl H2O , and 12.5 µl DNase master mix.
Incubate at RT for 10min.
3. Purification of RNA using Zymo RNA clean up and concentrator kit:
Follow the manufacturer's protocol, adding 100 µl RNA binding buffer and 300 µl ethanol as the initial step before transferring to column
Elute in 15 µl water. 

Note: Can also use RNeasy MinElute kit, using manufacturer's protocol
4. Reverse transcription with iScript.
For each sample, mix:
1 µl purified RNA
3 µl 5x iScript reagent
11 µl H2O
(total volume = 15 µl)

Also set up one no-RT reaction, same as above but using the iScript no-RT control reagent instead.

Run RT reaction as per manufacturer's protocol:
25°C 5 min
46°C 20 min
95°C 1 min

Before proceeding to PCR, add 35 µl H2O to each reaction.
5. PCR with Taq polymerase and primers flanking the insertion/deletion/mutation
Mix 4 µl virus cDNA with 46 µl of PCR mix containing :
0.4 µl dNTP (25 mM)
5 µl 10x Standard Taq buffer
0.25 µl Taq (5 U/ µl)
2 µl Forward primer (10 µM)
2 µl Reverse primer (10 µM)
36.35 µl dH2O

Note: other RT and polymerase reagents should work just as well. A proofreading polymerase may be preferable if checking for point mutations
6. Analysis of fragments
Run fragments on agarose gel to check that you have obtained the expected sizes. If appropriate, gel purify the product and send for Sanger sequencing.
TITLE: cAMP Regulated Cl- Current Protocol
AUTHORS: Robert  Harvey, Shailesh Agarwal

[DESCRIPTION]
cAMP Regulated Cl- Current Protocol

[STEPS]
1. Membrane currents were recorded from isolated pig ventricular myocytes using the whole-cell configuration of the patch clamp technique.
2. Isolated cells were placed in a perfusion chamber (Warner Instruments) on the stage of an inverted microscope (Olympus IX71) and bathed in a K+-free extracellular solution containing (in mM) NaCl 140, CsCl 5.4, CaCl22.5, MgCl2 0.5, glucose 5.5, and HEPES 5 (pH 7.4), maintained at room temperature. Nisoldipine (1 µM) was added to the extracellular solution to block L-type Ca2+ channels.
3. Cells were patched using microelectrodes with resistances between 1 and 2 MΩ. Access resistance was compensated to obtain series resistance errors of less than 5 mV.
4. Cells were dialyzed with a K+-free microelectrode solution containing (in mM): Cs-glutamate 130, TEA-Cl 20, EGTA 5, MgATP 5, TrisGTP 0.06, and HEPES 5 (pH 7.2).
5. Whole cell currents were recorded under voltage-clamp conditions using a Multiclamp 700B voltage clamp amplifier, Digidata 1440A computer interface, and pClamp 11 data acquisition and analysis software (Molecular Devices). Data were lowpass filtered at 4 kHz, and sampled at 10 kHz.
6. The membrane potential was held at -30 mV. A 100 ms test pulse to +50 mV was used to measure the cAMP-regulated Cl‑ current.
7. The time course of changes in the amplitude of the Cl‑ current was monitored by measuring the amplitude of the current recorded during the test pulse to +50 mV applied once every 5 s. 
8. A stable baseline (about 5 minutes) was obtained before application of test drugs.
9. The Cl‑ current was defined as the cAMP-induced current obtained by subtracting the current recorded before application of test-drug(s) from those recorded after.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div>
<div>
<p>This protocol is from 'Flower Ca<sup>2+</sup> channel in CME and ADBE' of Yao CK et al.</p>
<div>
<div>
<p> </p>
<p>Please see the manuscript for the full method details.</p>
</div>
</div>
</div>
</div>

[BEFORE_START]
<p>You'll need: </p>
<p> </p>
<p><strong>0 mM Ca<sup>2+</sup> hemolymph-like (HL)-3 solution:</strong></p>
<ul>
<li>70 mM NaCl</li>
<li>5 mM KCl</li>
<li>10 mM MgCl<sub>2</sub></li>
<li>10 mM NaHCO<sub>3</sub></li>
<li>5 mM trehalose</li>
<li>5 mM HEPES (pH 7.2) </li>
<li>115 mM sucrose</li>
</ul>
<p> </p>
<p> </p>
<p><strong> 2 mM Ca<sup>2+</sup>/5 mM K<sup>+</sup>/7 mM glutamate solution:</strong></p>
<ul>
<li>70 mM NaCl</li>
<li>5 mM KCl</li>
<li>10 mM MgCl<sub>2</sub></li>
<li>10 mM NaHCO<sub>3</sub></li>
<li>5 mM trehalose</li>
<li>5 mM HEPES (pH 7.2)</li>
<li>115 mM sucrose</li>
<li>2 mM CaCl<sub>2</sub> </li>
<li>7 mM Monosodium glutamate</li>
</ul>
<p> </p>
<p><strong>4 μM Fluo-4 AM (Invitrogen)/100 mM Ca<sup>2+</sup>/HL-3 solution to be loaded with Fluo-4 AM dye</strong></p>
<p> </p>
<p> </p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Quick Emulsion PCR Extraction Protocol
AUTHORS: Vijay K Chaudhary, Vaishali Verma, Amita Gupta, Vijay K. Chaudhary

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Emulsion PCR (ePCR) is an important technique that permits amplification of DNA molecules in physically separated picoliter volume water-in-oil droplets, and thus avoids formation of unproductive chimera and other artifacts between similar DNA sequences.However, the recovery of ePCR products involves repeated extraction with hazardous organic solvents followed by purification using silica-based columns, making the overall process cumbersome.  In this report, we have described a ‘Quick ePCR extraction protocol’ for the purification of ePCR products, which directly employs silica-based DNA purification columns and the products purified using this method have been found to be compatible with gene cloning and NGS applications.  The method described here makes ePCR easy, safe and within the reach of every laboratory. </div></div>

[STEPS]
?. Vigourously vortex the tube for 1 min.
?. Transfer entire emulsion PCR mixture (100 µl + the overlaid mineral oil) to a fresh 1.5 ml microfuge tube and add 500 µl of QIA PB purification buffer (5 x volume) directly the tube. For larger ePCR mixture, appropriate volume of PB buffer can be added.
?. Load the entire suspension on the QIAquick column.  Centrifuge at 13,000g for 1 min at room temperature (25°C). Discard the flow-through. This step may be repeated if the suspension is more than 700 µl.
?. Centrifuge the suspension at 13,000g for 1 min at room temperature (25°C).
?. Add 700 µl of QIA PE buffer to the column. Centrifuge at 13,000g for 1 min at room temperature (25°C). Discard the flow-through.
?. Centrifuge at 13,000g for 2 min at room temperature (25°C) to remove excess ethanol.  Transfer the column to a fresh microfuge tube.
?. Add 40 µl of QIA EB buffer to the column and incubate at room temperature (25°C) for 5 mins. Centrifuge at 13,000g for 1 min at room temperature (25°C).
?. The eluted DNA is ready to use for downstream work and can be quantified using agarose gel electrophoresis or Nanodrop or any other method available with the user.
TITLE: Disposal of Environmental Samples from Water Bodies Infested with Zebra Mussels 
AUTHORS: Malcolm A Barnard, Stephen Powers

[DESCRIPTION]
Zebra Mussels (Dreissena polymorpha) are a highly invasive species that have been invading water bodies around the United States and beyond. This protocol for sample disposal for sample collected from know zebra mussel infested water bodies to prevent spread though local water treatment facilities by bleaching the samples to kill any larvae or other potential zebra mussels in the sample water.

[STEPS]
SECTION: Sample Killing
1. Pour all environmental sample water from bottles into a dish pan.
SECTION: Sample Killing
2. Save the bottles for acid washing
 
.
SECTION: Sample Killing
3. Add 50 mL of bleach per 1 L of sample water.
4. Let the bleached sample sit for a minimum of four hours if not until the next day.
SECTION: Sample Disposal
5. Dispose of bleached sample down drain with excess water or other waste pathway as described by local environmental health and safety guidelines.
SECTION: Sample Disposal
6. Save bin for future sample disposal or bleach work.
TITLE: Bridport Health Liver Support:-(SCAM EXPOSED 2022)
AUTHORS: bridporthealth  

[DESCRIPTION]
The body is built to withstand a lot of chaos, but most people take it for granted. Whether drinking alcohol serves as a social lubricant or a symptom of depression, it can lead to addiction for some people. With these years of imbibing, anyone who has spent time in a health class or Alcoholics Anonymous knows the damage that can ensue.

[STEPS]
TITLE: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
AUTHORS: Pietro La Vitola

[DESCRIPTION]
This protocol is designed for  choline acetyltransferase (ChAT) staining using the Millipore AB144P (RRID:AB_2079751) antibody. Tissue stained with this protocol include 30μm free-floating mouse brain sections. All tissue was from mice perfused with 4% PFA. 


[STEPS]
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
1. Wash tissue slices in tris-buffer saline (TBS: 0.05M Trizma base and 0.15M NaCl; pH: 7.6) for 10 minutes. Repeat x3.
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
4. Incubate in blocking buffer (5% donkey serum, 2% BSA, 0.5% triton X-100 in TBS) for 1 hour at room temperature.
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
6. Wash tissue slices in TBS-T (TBS+ 0.25% Triton x-100) for 10 minutes. Repeat x3.
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
7. Incubate in fluorescent secondary antibody at 1:200 dilution in 50% blocking buffer for 1 hour at room temperature.
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
8. Wash tissue slices in TBS-T(TBS+ 0.25% Triton X-100) for 10 minutes. Repeat x 2.
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
10. Mount free-floating sections on SuperFrost+ slides (if staining free-floating tissue) and let dry at room temperature for 15 minutes.
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
11. Coverslip with fluorescent mounting medium and #1.5 coverslips. Outline coverslip with clear nail polish and store at 4oC .
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
5. Incubate with  anti-ChAT primary antibody  (RRID:AB_2079751) at 1:500 dilution in 50% blocking solution for 48 hours at 4°C. 
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
2. Incubate in in 10% Methanol + 3% H2O2 in TBS for 20 min at room temperature
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
9. Wash tissue slices in TBS (0.05M Trizma base and 0.15M NaCl; pH: 7.6) for 10 minutes.
SECTION: ChAT Immunofluorescent Staining of medulla oblongata fixed sections
3. Wash tissue slices in tris-buffer saline (TBS: 0.05M Trizma base and 0.15M NaCl; pH: 7.6) for 10 minutes. Repeat x3.
TITLE: Forward Primer Reconstitution
AUTHORS: Allyson Hirsch, George  Testo

[DESCRIPTION]
Stock primers should not be directly used in a PCR because they are concentrated. Working from one tube is a bad idea, and thus it only takes a small amount of contamination to render your primers ineffective. For this reason, it is best practice to create working solutions that are of lower concentrations. The concentration of choice for a working primer solution is user-dependent. However, 100uM is used for this protocol.

[BEFORE_START]
Make sure you work in a DNA-free environment. Preferably in a PCR preparation hood. This is to avoid contamination of your stock and working primer solutions.

Use PCR-grade water (DNase- and RNase-free) to reconstitute and dilute your primers.

Use filter pipette tips to prevent contamination via pipetting.

[STEPS]
SECTION: Preparing Forward Primer
1. Vortex and spin down primer tube: this breaks up the solidified primer at the bottom of the tube and brings the primer debris up from the bottom of the tube.
SECTION: Preparing Forward Primer
2. Look for nM (nanoMoles) on the printed primer label and circle. 

Ex: 779 µL

SECTION: Preparing Forward Primer
3. Multiple nM by 10, then add that much PCR grade water to the tube.

Ex: 779 µL

SECTION: Incubating Reconstitution
4. Vortex new 100 micromolar (µM) tube.

SECTION: Incubating Reconstitution
5. Let sit for at least 15 min (optimum: 1-2 hours).
SECTION: Incubating Reconstitution
6. Label stock tube as “100uM” on cap.
SECTION: Incubating Reconstitution
7. Once the sitting period has elapsed, place new 100 micromolar (µM) stock in a -20 °C freezer.

SECTION: Incubating Reconstitution
7.1. For Working Concentration of Forward Primer:

100 µL of FP stock + 900 µL molecular grade H2O
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
In this protocol we show how to perform the taxonomic biomarker discovery operation using <a href="https://bitbucket.org/nsegata/lefse" target="_blank">LEfSe</a>.

[BEFORE_START]
REQUIREMENTS: <a href="https://bitbucket.org/nsegata/lefse" target="_blank">LEfSe</a> installed (and in the system path), and the <a href="http://matplotlib.org/" target="_blank">matplotlib</a> python library installed. Alternatively, the users are welcome to use the <a href="http://huttenhower.sph.harvard.edu/lefse" target="_blank">Galaxy interface for LEfSe</a>. LEfSe can be downloaded using using <a href="http://mercurial.selenic.com/" target="_blank">Mercurial</a>: <code>hg clone ssh://hg@bitbucket.org/nsegata/lefse</code> or using the direct links to the <a href="https://bitbucket.org/nsegata/lefse/get/default.zip" target="_blank">zip</a>, <a href="https://bitbucket.org/nsegata/lefse/get/default.tar.gz" target="_blank">gz</a>, or <a href="https://bitbucket.org/nsegata/lefse/get/default.tar.gz" target="_blank">bz2</a> archives. Notice that LEfSe has some additional <a href="https://bitbucket.org/nsegata/lefse/src/ee1653ca297801562a3118aa606d063085be20d8/requirements.txt?at=default" target="_blank">requirements</a>.

[GUIDELINES]
We first need to specify the conditions (or classes) used for the biomarker discovery. Examples could be host disease states from which a gut microbiome was sampled, or environmental conditions (e.g. pH, environmental contaminant) from which a microbial community was sampled. In this tutorial, we will use body sites (contrasting the tongue dorsum microbiome with the buccal mucosa microbiome).<br /><br />The commands reported in this protocol can be retrieved as a <a href="https://bitbucket.org/nsegata/metaphlan/wiki/4_lefse_biomarker_discovery.sh" target="_blank">bash script</a>.

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p> </p>
<p>Coffee is considered as a major source of dietary antioxidants; some are present in the green bean, whereas others are generated during roasting.  Coffee roasting, the process of the heating of green coffee beans transforming them into black coffee beans, transforms the chemical and biological properties of coffee beans.</p>
<p>The recent studies have reported that coffee is inversely associated with basal cell carcinoma and endometrial cancer type I. Coffee drinking has been also inversely related to colorectal cancer and liver cancer.<strong>  </strong>Regarding oral cancer, some studies reported an as­sociation of high coffee consumption to an augmented risk of oral cancer, while others showed a clear in­verse association with the risk of oral cancer.</p>
<p>The main objective of our study was to evaluate anti proliferative and apoptotic activity of green coffee, in comparison to roasted coffee and decaffeinated coffee on cultured OSCC cell line.  </p>

[STEPS]