A Modified Simple Method for Induction of Myocardial Infarction in Mice

  • Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging and require commercially available apparatus. The current study describes a simple and efficient process for induction of MI in mice with readily available materials.
  • The mouse chest skin was cut open under stable anesthesia with a simplified anesthesia device made of centrifuge tubes. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an Corning Centrifuge Tubes MI model can be established in mice within 2-3 min with readily available materials.

Extending the working properties of liquid platelet-rich fibrin using chemically modified PET tubes and the Bio-Cool device

 Centrifuge Tube
Centrifuge Tube
  1. Platelet-rich fibrin (PRF) has been utilized in regenerative medicine as a concentration of autologous platelets and growth factors that stimulates tissue regeneration. More recently, liquid-PRF (also called injectable-PRF; i-PRF) has been brought to market utilizing PET plastic tubes. Due to new advances made in tube technology, the first aim of the present study was to investigate the liquid consistency of liquid-PRF utilizing both standard and chemically modified PET plastic tubes. Furthermore, it is well known that the conversion of PRF into a fibrin matrix is derived from the temperature-controlled enzymatic process that converts liquid fibrinogen and thrombin to solid fibrin. This study also investigated for the first time the use of a cooling device (Bio-Cool) to extend the liquid working properties of liquid-PRF.
  2. In total, 30 participants enrolled in this study. From each patient, four tubes of liquid-PRF were drawn, two standard white Vacuette tubes and two blue chemically modified hydrophobic tubes. Following centrifugation at 700 RCF-max for 8 min in a Bio-PRF horizontal centrifuge, one white and one blue tube were kept upright at room temperature, while the other white and blue tube were placed within the cooling device. Thereafter, the liquid-PRF layers were monitored over time until clotting occurred. Patient gender, age, and altitude above sea level (+ 5000 ft) were recorded and compared for clotting times.
  3. The findings from the present study demonstrated that the chemically modified PET tubes performed 37% better than the control tubes (extended the working properties of liquid-PRF by over 20 min). Most surprisingly, tubes kept in the cooling device demonstrated an average of 90 min greater working time (270% improvement). While patients living at altitude did significantly improve the clotting ability of liquid-PRF, no differences were observed when comparing male vs female or younger vs older patients in liquid-PRF clotting times.
  4. Cooling of blood following centrifugation represented a 270% improvement in working properties of liquid-PRF. Optimization of liquid-PRF tubes utilizing chemically modified hydrophobic PET tubes also delayed the clotting process by 37%. Patient gender and age had little relevance on liquid-PRF.
  5. The present findings demonstrate for the first time that cooling of liquid-PRF is able to extend the working properties of liquid-PRF by over 90 min. Thus for clinicians performing longer clinical procedures, the cooling of blood may represent a viable strategy to improve the working time of liquid-PRF in clinical practice. Bio-PRF; Cooling device; PRF; Platelet-rich fibrin; Wound healing.

Fixation and Immunostaining of Endogenous Proteins or Post-translational Modificationsin Caenorhabditis elegant.

  • Although the advent of genetically-encoded fluorescent markers, such as the green fluorescent protein (GFP; Chalfie et al., 1994 ), has enabled convenient visualization of gene expression in vivo, this method is generally not effective for detecting post-translational modifications because they are not translated from DNA sequences.
  • Genetically-encoded, fluorescently-tagged transgene products can also be misleading for observing expression patterns because transgenes may lack endogenous regulatory DNA elements needed for precise regulation of expression that could result in over or under expression. Fluorescently-tagged proteins created by CRISPR genome editing are less prone to defective expression patterns because the loci retain endogenous DNA elements that regulate their transcription (Nance and Frøkjær-Jensen, 2019).
  • However, even CRISPR alleles encoding heritable fluorescently-tagged protein markers can result in defects in function or localization of the gene product if the fluorescent tag obstructs or otherwise interferes with important protein interaction domains or affects the protein structure. Indirect immunofluorescence is a method for detecting endogenous gene expression or post-translational modifications without the need for transgenesis or genome editing.
  • Here, we present a reliable protocol in which C. elegans nematodes are fixed, preserved, and permeabilized for staining with a primary antibody to bind proteins or post-translational modifications, which are then labeled with a secondary antibody conjugated to a fluorescent dye.
  • Use of this method may be limited by the availability of (or ability to generate) a primary antibody that binds the epitope of interest in fixed animals. Thousands of animals are simultaneously subjected to a series of chemical treatments and washes in a single centrifuge tube, allowing large numbers of identically-treated stained animals to be examined.
  • We have successfully used this protocol (O’ Hagan et al., 2011 and 2017; Power et al., 2020 ) to preserve and detect post-translational modifications of tubulin in C. elegans ciliated sensory neurons and to detect non-modified endogenous protein (Topalidou and Chalfie, 2011).

Transfer some Hydrogel Containing 5-Fluorouracil and Etodolac Combination for Synergistic Oral Cancer Treatment

  • Oral cancer is one of the most common malignancies with an increased rate of incidence. 5-Fluorouracil (5FU) is an effective chemotherapeutic indicated for oral cancer treatment. Etodolac (Et), a cyclooxygenase-2 inhibitor, can be used as an adjuvant agent to sensitize cancer cells to chemotherapy. The aim of this work was to prepare and characterize 5FU and Et dual drug-loaded transfersomes to treat oral cancer.
  • Transfersomes were prepared by thin-film hydration method and characterized for the average particle size and zeta-potential using dynamic light scattering and scanning electron microscopy techniques.
  • The prepared transfersomes were further characterized for their drug loading, entrapment efficiencies using amicon centrifuge tubes and drug release behavior using cellulose membrane. The synergistic activity of dual drug-loaded transfersomes was studied in FaDu oral cancer cells. Results showed that the average particle size, polydispersity index, and zeta potential were 91±6.4 nm, 0.28±0.03, and (-)46.9±9.5 mV, respectively, for 5FU- and Et (1:1)-loaded transfersomes.

Centrifuge Tubes, 50 ml , Black ,PP,Sterile

LC8008-025 GenDepot 25/Bag

Centrifuge Tubes, 50 ml , Black ,PP,Sterile

LC8008-500 GenDepot 500/CS

Centrifuge Tubes, 15 ml , Black ,PP,Sterile

LC8007-050 GenDepot 50/Bag

Centrifuge Tubes, 15 ml , Black ,PP,Sterile

LC8007-500 GenDepot 500/CS

Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Bag

LC8005-500 GenDepot 500/CS

Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Bag

LC8003-500 GenDepot 500/CS

Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap,Self Standing, in Bag

LC8006-500 GenDepot 500/CS

Test Tube rack for 15 x 50 ml tubes

 -B2000-4-T500 Benchmark Scientific 1 PC

Centrifuge Tubes, 50 ml ,PP,Sterile,Plug-Seal Cap, in Rack

LC8004-500 GenDepot 500/CS

Centrifuge Tubes, 15 ml ,PP,Sterile,Plug-Seal Cap, in Rack

LC8002-500 GenDepot 500/CS

COMBI TUBE HOLDER, 6X15/50 ML TUBES

480148 CORNING 1/pk

ERLENMEYER FLASK, 50 ML, PP

4985P-50 CORNING 6/pk

ERLENMEYER FLASK, 50 ML, PMP

4990P-50 CORNING 6/pk

Block, 15 x1.5ml centrifuge tubes

BCM1405-B15 Bio Basic 1 pcs, 1 UNIT

500 ML DMEM (DULBECCO'S MODIFICATION OF EAGLE'S MEDIUM)/F12 50:50 MIX WITHOUT L-GLUTAMINE

15-090-CV CORNING 500 mL/pk

0.2 ml 8-Strip Tubes, 120 Strips of 8 Tubes

GP301 GenDepot 960 tubes

0.2 ml 8-Strip Tubes, 250 Strips of 8 Tubes

GP302 GenDepot 2,000 tubes

0.2 ml 12-Strip Tubes, 80 Strips of 12 Tubes

GP305 GenDepot 960 tubes

Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 1 mL

NGL40-B001 BioAssayWorks 1 mL

Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 9 mL

NGL40-B009 BioAssayWorks 9 mL

Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 44 mL

NGL40-B044 BioAssayWorks 44 mL

Naked Gold 40 nm Concentrated Gold Sol, 50 O.D./mL, 100 mL

NGL40-B100 BioAssayWorks 100 mL

Test Tube rack for 30 x 50 ml tubes

  B2000-8-T500 Benchmark Scientific 1 PC

Test Tube rack for 40 x 0.5 ml tubes

B2000-4-T5 Benchmark Scientific 1 PC

FALCON® 15 ML POLYSTYRENE CENTRIFUGE TUBE, CONICAL BOTTOM, WITH DOME-SEAL SCREW CAP, STERILE, 50/BAG, 500/CASE

352095 CORNING 50/pk

0.2 ml 8-Strip Tubes & Flat Caps, 250 Strips of 8 Tubes and Caps

GP203 GenDepot 2000 tubes/caps

0.2 ml 8-Strip Tubes & Domed Caps, 120 Strips of 8 Tubes and Caps

GP204 GenDepot 960 tubes/caps

0.2 ml 8-Strip Tubes & Domed Caps, 250 Strips of 8 Tubes and Caps

GP205 GenDepot 2,000 tubes/caps

0.3 ml 8-Strip Tubes & Domed Caps, 250 Strips of 8 Tubes and Caps

GP209 GenDepot 2,000 tubes/caps

0.2 ml 12-Strip Tubes & Flat Caps, 80 Strips of 12 Tubes and Caps

GP210 GenDepot 960 tubes/caps

0.2 ml 12-Strip Tubes & Domed Caps, 80 Strips of 12 Tubes and Caps

GP211 GenDepot 960 tubes/caps

0.2 ml 8-Strip Tubes & Flat Caps, 125 Strips of 8 Tubes and Caps

GP903 GenDepot 1000 tubes/caps

0.2 ml 8-Strip Tubes & Dome Caps, 125 Strips of 8 Tubes and Caps

GP904 GenDepot 1000 tubes/caps

Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 1 mL

NG40-B001 BioAssayWorks 1 mL

Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 9 mL

NG40-B009 BioAssayWorks 9 mL

Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 44 mL

NG40-B044 BioAssayWorks 44 mL

Naked Gold 40 nm Concentrated Gold Sol, 15 O.D./mL, 100 mL

NG40-B100 BioAssayWorks 100 mL

MINI BIOREACTOR, 50 ML CENTRIFUGE TUBE WITH VENT CAP, STERILE

431720 CORNING 25/pk

Test Tube rack for 41 x 15 ml tubes

  B2000-4-T150 Benchmark Scientific 1 PC

Test Tube rack for 76 x 15 ml tubes

  B2000-8-T150 Benchmark Scientific 1 PC

50 ml Reagent Reservoir

GR238016 Genorise Scientific Pack of 200

50ml Conical Centrifuge Tubes, Green Cap, 25/Bag

CT788-G Bio Basic 1PK, 25UNIT

50ml Conical Centrifuge Tubes, Yellow Cap, 25/Bag

CT788-Y Bio Basic 1PK, 25UNIT

0.2 ml 8-Strip Tubes & Optically Clear Caps, 120 Strips of 8 Tubes and Caps

GP207 GenDepot 960 tubes/caps

Test Tube rack for 40 x 1.5/2.0 ml tubes

  B2000-4-T1520 Benchmark Scientific 1 PC

ML 228

A4508-50 ApexBio 50 mg

ML-204

9448-50 Biovision

ML 213

B5628-50 ApexBio 50 mg

ML 171

B5672-50 ApexBio 50 mg

ML 204

B5696-50 ApexBio 50 mg

ML SA1

B5700-50 ApexBio 50 mg

ML 190

B5741-50 ApexBio 50 mg

ML 337

B5786-50 ApexBio 50 mg

ML 145

B7604-50 ApexBio 50 mg

ML 154

B7765-50 ApexBio 50 mg

ML 348

B4933-50 ApexBio 50 mg

ML-090

C4746-50 ApexBio 50 mg

ML-335

C4792-50 ApexBio 50 mg

ML-297

C5045-50 ApexBio 50 mg

0.2 ml 8-Strip Low Profile Tubes, 120 Strips of 8 Tubes

GP303 GenDepot 960 tubes

0.2 ml 12-Strip Low Profile Tubes, 80 Strips of 12 Tubes

GP304 GenDepot 960 tubes

0.2 ml 8-Strip Low-Profile Tubes & Flat Caps, 120 Strips of 8 Tubes and Caps

GP201 GenDepot 960 tubes/caps

0.2 ml 8-Strip Low-Profile Tubes & Domed Caps, 120 Strips of 8 Tubes and Caps

GP202 GenDepot 960 tubes/caps

0.2 ml 8-Strip Tubes & Domed Caps, Assorted Colors, 250 Strips of 8 Tubes and Caps

GP206 GenDepot 2,000 tubes/caps

0.2 ml 8-Strip White Tubes & Optically Clear Caps, 120 Strips of 8 Tubes and Caps

GP208 GenDepot 960 tubes/caps

AXYPREP MAG DYECLEAN KIT- 50 ML - 5000 PREPS

MAG-DYECL-50 CORNING 1/pk

RESERVOIR, POLYPROPYLENE, 50 ML, V-BOTTOM, NONSTERILE, BULK

RES-V-50 CORNING 100/pk

50 ML AMPHOTERICIN B, LIQUID 250 µG/ML SOLUBILIZED

30-003-CF CORNING 50 mL/pk

10 ML GENTAMICIN SULFATE, LIQUID 50 MG/ML SOLUTION

30-005-CR CORNING 10 mL/pk

50 ML KANAMYCIN SULFATE, LIQUID, 5,000 µG/ML SOLUTION

30-006-CF CORNING 50 mL/pk

Dressed Gold Streptavidin at 50 O.D./mL, 1 mL

DSTAL-B001 BioAssayWorks 1 mL

Dressed Gold Streptavidin at 50 O.D./mL, 9 mL

DSTAL-B009 BioAssayWorks 9 mL

Dressed Gold NeutrAvidin at 50 O.D./mL, 1 mL

DNTAL-B001 BioAssayWorks 1 mL

Dressed Gold NeutrAvidin at 50 O.D./mL, 9 mL

DNTAL-B009 BioAssayWorks 9 mL

Antigen Retrieval Solution, Citrate pH 6.0 (50 ml, 100X)

ARS6-50 Alpha Diagnostics 5 L

Antigen Retrieval Solution, EDTA pH 8.0 (50 ml, 100X)

ARS8-50 Alpha Diagnostics 5 L

Chloramphenicol Solution (50 mg/ml)

C090-10ML TOKU-E 10 mL

Chloramphenicol Solution (50 mg/ml)

C090-10x10ML TOKU-E 10 x 10 mL

BrightGreen miRNA qPCR MasterMix-Low ROX

MasterMix-mL ABM 4 x 1.25 ml for 500 reactions

Rat Monoclonal Anti-Mouse CD4 Pur. Low Endotoxin (Clone YTS 191.1) (rat IgG2b)

MCD004-ML Alpha Diagnostics 100 ug

Rat Monoclonal Anti-Mouse CD25 , Pur. Low Endotoxin (Clone PC61.5.3) (rat IgG1)

MCD025-ML Alpha Diagnostics 100 ug

ML-7 hydrochloride

A3626-50 ApexBio 50 mg

ML 218 hydrochloride

B5622-50 ApexBio 50 mg

ML 120B dihydrochloride

B5756-50 ApexBio 50 mg

ML 9 hydrochloride

B6300-50 ApexBio 50 mg

ML 10302 hydrochloride

B7413-50 ApexBio 50 mg

ML 00253764 hydrochloride

B7715-50 ApexBio 50 mg

AXYPREP FRAGMENTSELECT-I KIT- 50 ML - 380 PREPS

MAG-FRAG-I-50 CORNING 1/pk

RESERVOIR, POLYPROPYLENE, 50 ML, V-BOTTOM, STERILE, BULK

RES-V-50-S CORNING 5/pk

Dressed Gold Streptavidin at 15 O.D./mL, 1 mL

DSTA-B001 BioAssayWorks 1 mL

Dressed Gold Streptavidin at 15 O.D./mL, 9 mL

DSTA-B009 BioAssayWorks 9 mL

Dressed Gold NeutrAvidin at 15 O.D./mL, 1 mL

DNTA-B001 BioAssayWorks 1 mL

Dressed Gold NeutrAvidin at 15 O.D./mL, 9 mL

DNTA-B009 BioAssayWorks 9 mL

Antigen Retrieval Solution, Tris-EDTA pH 9.0 (50 ml, 100X)

ARS9-50 Alpha Diagnostics 5 L

Dressed Gold Protein A at 50 O.D./mL, 1 mL

DPRAL-B001 BioAssayWorks 1 mL

Dressed Gold Protein A at 50 O.D./mL, 9 mL

DPRAL-B009 BioAssayWorks 9 mL

Dressed Gold Protein G at 50 O.D./mL, 1 mL

DPRGL-B001 BioAssayWorks 1 mL

Dressed Gold Protein G at 50 O.D./mL, 9 mL

DPRGL-B009 BioAssayWorks 9 mL

Dressed Gold Protein L at 50 O.D./mL, 1 mL

DPRLL-B001 BioAssayWorks 1 mL

Dressed Gold Protein L at 50 O.D./mL, 9 mL

DPRLL-B009 BioAssayWorks 9 mL
The highest encapsulation efficiency achieved was 36.9±3.8% and 79.8±6.4% for 5FU and Et (1:1), respectively. Growth inhibition studies in FaDu cells using different concentrations of 5FU and Et showed a combination index of 0.36, indicating a synergistic effect. The FaDu cell uptake of drug-loaded transfersomes was significantly (p<0.05) greater than that of free drugs. The transfersome hydrogel made of HPMC (2% w/w) showed similar flux, lag time, and permeation coefficient as that of drug-loaded transfersomes across excised porcine buccal tissue. In conclusion, 5FU and Et transfersome hydrogel can be developed for localized delivery to treat oral cancer.

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