eamag/protocols-with-steps huggingface dataset:
TITLE: Sea Cucumber Sperm Extraction from Gonads
AUTHORS: Jon Eilers

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">How to extract sperm and gonadal tissue from sea cucumber gonads</div></div>

[STEPS]
?. Secure mesh (panty hose or other mesh like material) to top of funnel and place funnel into a falcon tube
?. Place gonads onto mesh and begin gently mashing with surgical spatula
An opaque fluid should begin accumulating in the bottom of the tube
TITLE: Crystallisation of Zika NS5 RdRp
AUTHORS: Anu V. Chandran, Peter Marples, Martin austin Walsh

[DESCRIPTION]
The main aim of this work was to identify small molecules that bind Zika NS5 RdRp (catalytic RNA-dependent RNA polymerase domain) through X-ray fragment-based screening. The Zika NS5 RDRP domain was cloned, expressed, purified, and crystallised.  Suitable crystals for fragment screening were produced and optimised allowing an extensive fragment campaign to be performed. A native high-resolution structure was determined at 1.8Å and formed the basis for the fragment campaign.  

[STEPS]
SECTION: Crystallisation experiment
3. Protein and buffer requirements:
28.8 µL5 mg/mL  
3.264 mL 
SECTION: Crystallisation experiment
5. Dispense 34 µL into SwissCI 3 lens plate reservoir wells using a 100 µl multi-channel pipette.
Dispense 100 nL5 mg/mL    to each lens using the SPT mosquito.
Dispense  50 nL to each lens using the SPT mosquito.

Drop ratio: 2:1 ratio (100 nl   : 50 nl reservoir solution)
Final drop volume: 150 nl
SECTION: Crystallisation experiment
4. Crystallisation screen composition:
Morpheous I E10 condition
 
0.12 Molarity (M) Ethylene glycols
0.1 Molarity (M) Buffer system 3 pH 8.5
30 % v/v Precipitant Mix 2
Stock solutions used:
Morpheous I E10 condition

 

SECTION: Crystallisation experiment
7. Crystal form after ~24 h.
 


SECTION: Crystallisation experiment
6. Incubate at 20 °C for 1440 min in Formulatrix Rock Imager.

Imaging Schedule: The first images are taken after 12 h and the imaging schedule follows a Fibonacci sequence of days for further collections. 
SECTION: Equipment needed
2. Formulatrix Rock Imager (or incubator of choice)
SPT mosquito
 
P100 8 multi-channel pipette
SwissCI 3 lens plate

SECTION: Data collection at Synchrotron
8. Diamond Light Source
Unattended Data Collection (UDC)
Data Collection Temperature: 100K
Detector: DECTRIS EIGER2 X 9M
Beamline: I04-1
Wavelength:  0.9212 Å
Resolution (Å): 1.78
Beam Size (μm): 60 X 50
Number of images: 3600
Oscillation: 0.10°
Exposure (s): 0.0020
Transmission (%): 100
Flux (ph/s): 9.50e+11
SECTION: Zika NS5 RdRp expression and purification
1. The protein used for crystallisation was expressed and purified using the following protocol.
TITLE: Sampling strategies and sample preservation Acropora
AUTHORS: Iliana Baums

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Protocol describes sampling strategies, perservation and shipping of Acropora samples for </div><div class = "text-block">Applied Biosystems™ Axiom™ Coral Genotyping Array – 550961 (384f)</div><div class = "text-block">Applied Biosystems™ Axiom™ Coral Genotyping Array – 550962 (Mini 96)</div></div>

[STEPS]
TITLE: JAX - 10x Genomics - NEXTGEM Multiome ATAC + Gene Expression
AUTHORS: William F F F. Flynn

[DESCRIPTION]
Standard 10x Genomics - NEXTGEM Multiome protocol used by the JAX Single Cell Biology lab for the simultaneous single nucleus chromatin accessibility and gene expression assay.

[STEPS]
1. Follow the 10x Genomics NEXTGEM Single Cell Multiome ATAC+Gene expression protocol (CG00338).
TITLE:  Attention! NextGen Pharma Keto Reviews : Scam Or Legit  | Must Read! 
AUTHORS: Warapen  Treekitkarnmongkol

[DESCRIPTION]
The  easiest way to start losing weight today is to visit the  official NextGen Pharma Keto website and grab a bottle for yourself. The  company offers some discounts to people who purchase the goods on their  website.

[STEPS]
TITLE: Phenol-chloroform extraction and ethanol precipitation of RNA
AUTHORS: Tomasz Suchan

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Based on the protocol form NEB HiScribeTM T7 High Yield RNA Synthesis Kit</div></div>

[STEPS]
?. Adjust the reaction volume to 180 μl by adding 160 μl nuclease-free water. Add 20 μl of 3 M sodium acetate, pH 5.2 or 20 μl of 5 M ammonium acetate (1/10 of the total volume), mix thoroughly.
?. Extract with an equal volume (200 μl) of 1:1 phenol/chloroform mixture. Vortex and centrufugate at maximum speed for 5 min.
?. Precipitate the RNA by adding 2 volumes of -20°C 100% ethanol. Incubate at –20°C or on dry ice for at least 30 minutes.
?. Remove the supernatant and rinse the pellet with 500 μl of cold 70% ethanol.
?. Resuspend the RNA in 50 μl of 0.1 mM EDTA. Store the RNA at –20°C or below.
?. Collect the aqueous (upper) phase and transfer to a new tube.
?. Repeat steps 2 and 3.
?. Collect the pellet by centrifugation at 4°C for 30 min.
?. Quick spin and collect the last drop of ethanol.
?. Dry the sample.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><span style="color: #3c3c39;">Bioinformatic pipleine used to process all sequence reads. </span></p>

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

[DESCRIPTION]
For use in "<a href="https://www.protocols.io/view/Isolation-of-cyanophages-by-liquid-enrichment-assa-dp65rd" target="_blank">Isolation of cyanophages by liquid enrichment assay</a>"

[STEPS]
?. 
?. 
?. 
?. 
?. 
?.
TITLE: SEM gold coat protocol
AUTHORS: Elizabeth Fozo

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Protocol for how to the gold coater for SEM samples</div></div>

[STEPS]
?. [This coater is in the lab sample prep room in JIAM.]
Lift glass unit – hold it together and set it to the side.
?. [This coater is in the lab sample prep room in JIAM.]
Add samples on the silicon chip and mounted post.
?. [This coater is in the lab sample prep room in JIAM.]
Turn pump on (switch on the left).
?. [This coater is in the lab sample prep room in JIAM.]
Open cylinder valve on left.
?. [This coater is in the lab sample prep room in JIAM.]
Pump for a couple of minutes.
?. [This coater is in the lab sample prep room in JIAM.]
Gas leak – bleed argon for 2 – 3 cycles.
?. [This coater is in the lab sample prep room in JIAM.]
Push and hold test.
?. [This coater is in the lab sample prep room in JIAM.]
Open gas leak valve – go fast, but not too fast.
?. [This coater is in the lab sample prep room in JIAM.]
Bring the number to around 18 then hit the start button.
TITLE: Virus Production and Administration
AUTHORS: Sarkis  Mazmanian, Bryan  Yoo, Jessica  Griffiths

[DESCRIPTION]
Protocol for virus production and administration to mice in Yoo et al 2021

[STEPS]
SECTION: Virus Production
4. Virus is precipitated from cells and supernatant with an 8% PEG solution (wt/vol), and purified by ultracentrifugation using 15%, 25%, 40%, 60% stacked iodixanol gradients.
SECTION: Virus Production
3.  Briefly, human embryonic kidney (HEK293T) cells were triple-transfected with pUCmini-iCAP-AAV-PHP.S, pHelper Plasmid, and one of the following pAAV genomes (CAG-NLS-GFP, hSYN1-tDTomato, GFAP-tDTomato, hSYN1-mRuby2, hSYN1-DiO-mRuby2, hSYN1-mNeonGreen, hSYN1-DiO-mNeonGreen, hSYN1-mTurquoise2, hSYN1-DiO-mTurquoise2, hSYN1-DiO-hM3Dq-mRuby2, CAG-GCaMP6F) in a DPBS + polyethylenimine (PEI)
SECTION: Virus Production
2. Cells were grown in DMEM + Glutamax + Pyruvate (Gibco, Gaithersburg, MD-Stock# 10569-010) + 5% FBS + non-essential amino acids (Gibco, Gaithersburg, MD-Stock# 11140-050), penicillin-streptomycin (Gibco, Gaithersburg, MD-Stock# 15070-063). 
SECTION: Virus Production
1. Virus was produced using methods described in Challis et. al. (Challis et al., 2019) 
SECTION: Virus Administration
5. Virus was titered to 1012 viral genomes (vgs) and resuspended to a volume of 100µl with sterile PBS.
SECTION: Virus Administration
6. Mice were anesthetized using 2% isoflurane, and virus was injected retro-orbitally.
TITLE: 10% Tween 20
AUTHORS: Allen Institute for Brain Science

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol is used to prepare 10% Tween 20. Tween 20, when added to aqueous solutions acts as a surfactant, decreasing the surface tension of the solution. It also punches holes in membranes, allowing easier penetration of macromolecules into cells and organelles, and reduces non-specific binding of antisera in immunohistochemistry.</div><div class = "text-block"><span style = "font-weight:bold;">Note</span><span>: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.</span></div></div>

[STEPS]
TITLE: Mito-Keima assay to assess mitophagy
AUTHORS: Thanh Ngoc Nguyen

[DESCRIPTION]
This protocol details the procedure of mito-keima assay to assess mitophagy.

[STEPS]
SECTION: Procedure
1. Seed the HeLa cells the day before the treatment day in 24 well plates.
 

SECTION: Procedure
2. The next day, make sure the seeded cells are spreading out.
 

SECTION: Procedure
3. Aspirate off the old media and treat each well with 0.5 mL of growth media containing 4 micromolar (µM) Antimycin A, 10 micromolar (µM) Oligomycin and 10 micromolar (µM) QVD for indicated times.
 

SECTION: Procedure
4. 2 hours prior to harvesting, feed the untreated wells with 1 mL of warm growth media.
SECTION: Procedure
5. Aftrer treatment, harvest the cells by trypsinisation.
SECTION: Procedure
6. Resuspend the cell pellets in 150 µL of FACS buffer, transfer to pre-chilled FACS tubes and keep the samples on ice in the dark for analysis.

SECTION: Procedure
3.1. Treat the longest time points first.
SECTION: Procedure
5.1. Pre-chill eppies on ice.

SECTION: Procedure
5.2. Aspirate the media thoroughly from the wells.
SECTION: Procedure
5.4. Aspirate 1x PBS and add 150 µL of trypsin and incubate at 37 °C for 2 min-5 min.
 

SECTION: Procedure
5.5. Add 300 µL of growth media to each well.
SECTION: Procedure
5.7. Centrifuge the eppies at 1000 x g for 2 min at 4 °C.
SECTION: Procedure
5.3. Wash the wells with 0.5 mL of 1x PBS.
 

SECTION: Procedure
5.6. Mix well with a P1000 and transfer the cells to cold eppies.
SECTION: Procedure
5.8. Aspirate off the liquid.
SECTION: Procedure
1.1. Control cells include unstained cells (without any fluorescence proteins), cells that only express GFP-OPTN or GFP-NDP52 (to set up compensation so that GFP signal doesn’t bleach into mtKeima signal.
TITLE: Determining Lipid Content in Embryos using Nile Red Fluorescence
AUTHORS: Luciano Bonilla, Peter J. Hansen

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The following protocol is derived from the protocol described by Genicot et al. (Theriogenology 63: 1181- 1194, 2005) and is based on the fluorescence emitted by Nile Red when in association with lipid (see Fowler and Greenspan, J Histochem Cytochem 33: 833-836, 1985). </div></div>

[STEPS]
TITLE: Propagating Serratia phage 92A1 and host
AUTHORS: Adair Borges

[DESCRIPTION]
This is a protocol for propagating Serratia phage 92A1 and its host, Serratia strain 92. Serratia phage 92A1 (Genbank OR088902.1) is a 174 kb, T4-like phage with a probable arabinose-based modification of its genome. You can use this protocol to generate large quantities of phage for DNA extraction and nucleoside chemical analysis.

[STEPS]
SECTION: Propagating Serratia strain 92
1. Streak culture out on an LB agar plate and incubate at room temperature (25 °C) for two days. You should see the formation of light tan colonies that have a slightly gooey appearance.

 

SECTION: Propagating Serratia strain 92
2. Set up a liquid culture of the host by inoculating 1 mL of LB broth with a single colony and incubating in a shaker at room temperature (25 °C) shaking at 200 rpm. Grow for two days. 

 


SECTION: Propagating phage 92A1
3. Infect 200 μl of host liquid culture with 10 μl of phage in a microcentrifuge tube. Let incubate on benchtop for 10 minutes.

 

SECTION: Propagating phage 92A1
4. Move 3 mL of molten 0.25% top agar (see recipe below) into a glass culture tube. Once the top agar is warm but not so hot that it will kill your cells (aim for lower than ~45 °C), add the pre-mixed phage and host to the tube.

 

SECTION: Propagating phage 92A1
5. Rapidly pour the inoculated top agar onto a room-temperature LB plate, and swirl to distribute a thin layer of top agar across the surface of the plate. Put it down and let it fully solidify. Do not disturb the plate until it is fully solidified. 
SECTION: Propagating phage 92A1
6. Incubate at room temperature (25 °C) for one day. You should see small dark plaques form, and if the input concentration of phage is correct, these plaques should merge to form a confluent zone of lysis.
SECTION: Propagating phage 92A1
7. To harvest the phage, flood the plate with 3 mL of SM buffer (recipe below) and incubate on benchtop for 10 minutes. This allows the phage to diffuse out of the top agar into the buffer layer, which can then be harvested.

 

SECTION: Propagating phage 92A1
8. Pipette the SM buffer with phage into a 15 mL conical tube, and add 300 μl of chloroform to the mixture. If you are combining multiple plates worth of phage, scale up the amount of chloroform you are using (300 μl for every 3 mL buffer).
SECTION: Propagating phage 92A1
9. Mix buffer and chloroform on a rocking shaker for 10 minutes. Use Parafilm to seal the top to make sure nothing leaks out.
SECTION: Propagating phage 92A1
10. Spin down at 15,000 × g for 5 min. You should see the cells and other debris collect in a cohesive opaque layer above the chloroform.
SECTION: Propagating phage 92A1
11. Collect the supernatant. If it's still cloudy, repeat the chloroform treatment and spin. Keep doing this until the supernatant is clear. 
SECTION: Propagating phage 92A1
12. Store the supernatant on 50 μl of chloroform in the fridge at 4 °C. This phage appears to be stable long term at 4 °C, but consider checking it every 6–9 months to make sure it's viable, and propagate it if you are losing titer.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><strong>Background </strong>Elderly patients undergoing hip fracture surgery (HFS) are at increased risk of postoperative venous thromboembolism (VTE). Therefore, combined postoperative mechanical and chemical thromboprophylaxis has been routinely performed after HFS in these patients. This <u>retrospective case-control study was conducted</u> to evaluate the additional effectiveness of preoperative mechanical thromboprophylaxis on the prevention of VTE following HFS in elderly patients.</p>
<p><strong>Methods</strong> Of 539 consecutive patients aged 70 years or older undergoing HFS, 404 (control group) did not receive preoperative mechanical thromboprophylaxis while 135 (study group) received mechanical thromboprophylaxis using an intermittent pneumatic compression device and graduated compression stockings from the time of admission until surgery. All patients received combined postoperative mechanical and chemical thromboprophylaxis following HFS in accordance with the same protocol. The incidence of symptomatic VTE confirmed by clinical symptoms and 3-dimensional CT angiography within one month of surgery was investigated in both groups.</p>
<p><strong>Results</strong> American Society of Anesthesiologists grade was higher (p=0.016) in the study group and more patients in this group had concomitant cardiovascular and neurologic diseases (p=0.005 and p=0.009, respectively). Meanwhile, more patients in the study group had received anticoagulant medication preinjury owing to comorbidities (39% vs 28%, p=0.025). The overall incidences of symptomatic deep vein thrombosis (DVT) and pulmonary embolism (PE) were 7.4% and 3.7% in the control group and 2.2% and 1.5% in the study group, respectively. On multiple logistic regression, symptomatic DVT significantly reduced in the study group (OR 0.28, p=0.042), meanwhile there was no significant difference in the incidence of symptomatic PE between the two groups (p=0.223).</p>
<p><strong>Conclusions</strong> Preoperative mechanical thromboprophylaxis may confer an additional benefit by preventing postoperative VTE without adding more risk of perioperative bleeding in elderly patients with hip fracture.</p>

[STEPS]
?.
TITLE: Statistical Considerations (Part 9 of Phase 3 study of Vaccine Candidate for COVID-19)
AUTHORS: Chris Ockenhouse, Chris Gast, Renee Holt, Jorge Flores

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>This is Part 9 of "Phase 3 randomized, double-blinded, placebo-controlled trial to evaluate the safety, immunogenicity, and efficacy of </span><span style = "font-weight:bold;">Vaccine Candidate</span><span> against COVID-19 in adults > 18 years of age"</span></div><div class = "text-block">This generic Phase 3 protocol was developed by the PATH team with support of the Bill and Melinda Gates Foundation. The aim of the collection is to share recommended best practices in designing and implementing a Phase 3 study of a COVID-19 vaccine candidate. As Phase 3 trials of different Vaccine Candidates  proceed around the world, following the same protocols will ensure consistency and comparability of the Phase 3 trial results.</div><div class = "text-block"><span style = "font-weight:bold;">Please note</span><span> that this is an evolving document, to be versioned and updated, based on community feedback and new data.</span></div></div>

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This staining was employed to detect Fetuin-A  in paraffin sections (1 μm thickness) of formalin-fixed human brain tissue. For fetuin-A staining, we used a monoclonal IgG2a mouse-anti-human antibody (clone MAHS-1, dilution 0.1-0.5 µg/mL), raised against purified human fetuin-A in our laboratories. Antibodies were diluted in a 1% dilution of Bovine serum albumin (BSA) in phosphate-buffered saline (PBS) and were immediately applied to the re-hydrated sections. Bound antibody was detected using Dako REAL™ Detection System, which employs APAAP immunochemistry and fast red chromogenic substrate (Dako K5000, Glostrup, Denmark) following the manufacturers protocol. Counterstaining was employed with Mayer´s hematoxylin solution (Roth, T160.1, one minute). The slides were then washed in demineralized water and dehydrated in graded alcohol (concentrations from 70% to 100%). After placing in xylene the sections were mounted (Roth, T160.1) and covered using coverslips.</p>

[GUIDELINES]
<p>- use with APAAP (Alkaline Phosphatase - Anti-Alkaline Phosphatase) Kit (Dako K5000, Glostrup, Denmark)</p>
<p>- positive control: human liver</p>

[STEPS]
SECTION: Clear slides
?. 
?. 
?. 
?. 
SECTION: Antigen retrieval
?. 
?. 
SECTION: Immunostaining
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
SECTION: Dehydrate and mount slides
?. 
?. 
?. 
?.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>A real-time PCR for Parapoxvirus targeting the DNA polymerase. It is used to screen human samples where Parapoxvirus is suspected.</p>
<p>This protocol is based on the published RVSS assay by Das et al 2017. Oligonucleotides have been modified and a different PCR kit is used.</p>

[GUIDELINES]
<ul>
<li>If using a different brand or model of real-time thermocycler, check the concentration of ROX is adequate.</li>
<li>Method assumes the user is familiar with the thermocycler and software used to run the protocol.</li>
</ul>

[STEPS]
?. 
?. 
?. 
?.
TITLE: Synthesis of fluorinated neonicotinoids
AUTHORS: Andrii Kyrylchuk, Andriy Bezdudnyy, Denys Klukovskyi

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This is a general protocol for synthesis of fluorinated neonicotinoid analogues by interaction of amines with 2-chloro-5-(chloromethyl)pyridine.</div></div>

[STEPS]
?. Dissolve   of corresponding substituted aniline in  of anhydrous acetonitrile in round-bottom flask, add  () of  2-chloro-5-(chloromethyl)pyridine and  () of anhydrous potassium carbonate.
2
5 ml
0.324 g
1.9
0.828 g
6
?. Put a water-cooled backflow condenser on top of the flask and heat the flask under vigorous stirring until the solution starts to boil.
?. Continue refluxing for several hours. Monitor reaction progress via thin layer chromatography (TLC).
?. After the reaction is complete, filter off inorganic salts and rigorously wash the sediment with dichloromethane.
?. Combine all liquid fraction and remove the solvents by evaporation under reduced pressure.
?. Purify the residue by preparative  (TLC) on SiO2 (eluent EtOAc/hexane 1:2).
TITLE: OCT-Embedded Tissue Preparation
AUTHORS: Stephen Fisher, Marielena Grijalva, Rong Guo, sarahjoh, Hieu Nguyen, John Renz, Jean G Rosario, Steven Rudich, Brian Gregory, Junhyong Kim, Kate O'Neill

[DESCRIPTION]
This protocol describes embedding of tissue samples in optimal cutting temperature (OCT) compound (Lin & Lin, 2020).  Tissue embedded in OCT can then undergo frozen sectioning on a microtome-cryostat.  The advantage of OCT embedding is it requires minimal tissue processing and tissue does not need to undergo fixation prior to snap freezing.   OCT blocks should be stored in -80°C freezer and can yield thin tissue sections (< 5µm). Formation of bubbles in the OCT can distort and disrupt tissue.  To minimize bubble formation, we recommend storing OCT upside down in a small bucket between uses. When ready for use, uncap OCT tube and dispense slowly to prevent bubbles from forming.  Once tissue is in cassette surrounded by OCT, snap freezing is done.   Ensure you have a portable liquid nitrogen dewar partially filled with liquid nitrogen. Place a piece of Styrofoam atop the liquid nitrogen to prevent direct contact of the embedding molds with the cold liquid.

[STEPS]
1. Label biopsy molds appropriately.
2. Add a small amount of OCT to the bottom of each mold, ensuring no bubbles are present.
3. Place tissue piece in the center of the mold on top of the OCT layer.
4. Pour OCT slowly into the mold, ensuring the tissue piece is completely covered.
5. Using a small piece of aluminum foil, tightly wrap the mold. 

Tip: Label the outside of the aluminum for easy retrieval later.
6. Place foil-wrapped mold flat in a liquid nitrogen dewar on top of the Styrofoam.
7. After approximately one minute, move the frozen OCT mold to the dry ice container. 

Note: OCT will become opaque when frozen.
8. Store molds at -80℃ ultra-low freezer for Biobanking or until ready for downstream processing.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><strong>Background</strong>: VEGF is elevated in plasma of colon cancer patients and increases after surgery. This study's purpose was to determine plasma VEGF levels before, during, and after open and laparoscopic colorectal surgery performed for left colon cancer (CD).</p>
<p><strong>Patients and methods</strong>: a multicenter, prospective, observational, non-randomized study was planned to evaluate samples obtained preoperatively, intraoperatively, and on postoperative days (1, 3, 7, 15, 30). VEGF-A<sub>165</sub> values were determined via ELISA. Statistical analysis was a repeated Anova with interactions.</p>
<p><strong>Results</strong>: the recruitment finished in 4 years (71 patients). Baseline plasma VEGF-A<sub>165</sub> mean values (pVmv) resulted not significantly different in patients submitted to laparoscopy (125.4 pg/ml ± 47.7) and laparotomy (116.3 pg/ml ± 28.9). Intraoperatively, the highest difference was registered at the end of operations. In the 30-days postoperative period, the pVmv resulted significantly lower (G-G=0.45) in patients submitted to laparoscopy than in patients submitted to laparotomy (F= 144.6, p&lt;0.001), at time went on (F= 97.6, p&lt;0.001); the highest differences between groups were registered at day 7.</p>
<p><strong>Conclusions</strong>: the trial supports the hypothesis that cancer and surgical trauma are related to production of VEGF-A<sub>165</sub> and demonstrates that open surgery, in comparison to laparoscopy, is a stronger promoter of VEGF release.</p>

[GUIDELINES]
<p>No guidelines</p>

[STEPS]
?.