Bacteria
Julius P. Kreier in Infection, Resistance, and Immunity, 2022
Cholera remains a major problem in Third World countries and in any area where water supplies are contaminated with domestic sewage. Incidence of the disease increased about 20-fold between 1938 and 1988. The disease is caused by Vibrio cholerae, a curved Gram-negative rod that adheres to the mucosa within four to six hours. The cholera toxin of especially virulent strains (V. cholerae 01 or 0139) is an A-B subunit toxin (see E. coli toxins, above) that stimulates adenylate cyclase production which increases intracellular levels of cAMP. in turn, cAMP initiates secretion of both water and ions from the gut epithelial cells. It is the loss of water and salt that causes the shock characteristic of the disease. Ten to fifteen percent of those infected may die. Treatment is to restore water and electrolyte balance. The patient is made to consume sufficient balanced salt solution to replace the lost water and salt. Endemic infections occur in areas with poor sewage and water treatment facilities. Transmission occurs by contamination of food and water by the diarrheal feces. Between the epidemics the organisms probably survive as harmless parasites of copepods and other animals living in rivers, ponds and estuaries. It is probable that the organism came originally from rivers in northern India.
Indocyanine green and trypan blue in vitreoretinal surgery
A Peyman MD Gholam, A Meffert MD Stephen, D Conway MD FACS Mandi, Chiasson Trisha in Vitreoretinal Surgical Techniques, 2019
ICG stains the ILM and the lens capsule but not ERMs. A wide range of ICG concentrations (0.25–5 mg/ml) have been used for staining during vitrectomy.1 ICG powder should be dissolved with the aqueous solvent provided by the manufacturer; 10 ml of aqueous solvent is added to 25 mg ICG for a concentration of 2.5 mg/ml. After the solution has been shaken, it can be further diluted with balanced salt solution (BSS); 0.5 ml of this solution is diluted with 2 ml for a concentration of 0.5 mg/ml or is diluted with 0.5 ml of BSS for a concentration of 1.25 mg/ml.
Methods for in Vitro Skin Metabolism Studies
Francis N. Marzulli, Howard I. Maibach in Dermatotoxicology Methods: The Laboratory Worker’s Vade Mecum, 2019
Tissue culture media, such as minimal essential media (MEM), can be used to maintain skin viability in a flow-through diffusion cell. However, we have found that HEPES-buffered Hanks balanced salt solution (HHBSS) and Dulbecco modified phosphate-buffered saline are physiological buffers that also can be used (Collier et al., 1989). Vitamins and other cofactors in MEM are not required to maintain viability of skin in a diffusion cell for 24 h, and they sometimes can interfere in analytical techniques.
Dolutegravir potentiates platelet activation by a calcium-dependent, ionophore-like mechanism
Published in Journal of Immunotoxicology, 2022
Morris Madzime, Annette J. Theron, Ronald Anderson, Gregory R. Tintinger, Helen C. Steel, Pieter W. A. Meyer, Jan G. Nel, Charles Feldman, Theresa M. Rossouw
In order to determine the expression of CD62P on platelets, the method of Anderson et al. (2020) was followed. In brief, to PRP (20 μl) in a final volume of 1 ml Hanks’ balanced salt solution (HBSS, indicator-free, 1.25 mM calcium Ca2+, pH 7.4), either DMSO (control) or dolutegravir (at final concentrations of 5 and 10 µg/ml) were added. The tubes were then incubated for 10 min at 37 °C after which the platelets were activated with ADP (100 µM) for 5 min. Flow cytometry was then performed to determine the magnitude of expression of CD62P according to the proportion (%) of CD42a+ platelets expressing the adhesion molecule. In an additional series of experiments, the effects of dolutegravir, at a fixed concentration of 10 μg/ml, on the expression of CD62P by platelets activated with thrombin (0.6 NIH units/ml) or U46619 (5 µM) were investigated.
Association of Deepening of the Upper Eyelid Sulcus with the Incidence of Blepharoptosis after Glaucoma Filtration Surgery
Published in Seminars in Ophthalmology, 2020
Masaki Fukushima, Tatsuya Yunoki, Mitsuya Otsuka, Atsushi Hayashi
Glaucoma filtration surgery was performed by two glaucoma specialists (AH and MO). Subconjunctival and sub-Tenon’s anesthesia were performed using 2% lidocaine. When cataract surgery was performed simultaneously, the patients underwent phacoemulsification and intraocular lens implantation (PEA+IOL) by temporal corneal incision. The corneal incision was sutured with 10–0 nylon. An upper fornix-based conjunctival incision was made to expose the sclera. We created a 3.5 mm ×3.5 mm scleral flap and applied mitomycin C solution (MMC) (0.04 mg/mL) for 4 min. Then, MMC was sufficiently washed away with a balanced salt solution. When performing conventional trabeculectomy, sclerotomy was performed, and the trabecular meshwork was removed. Then peripheral iridectomy was performed. On the other hand, when using an Ex-PRESS® mini-glaucoma shunt device (Ex-Press; Alcon Laboratories, Fort Worth, TX, USA), a 25 G needle was inserted into the anterior chamber from the sclera–cornea transition zone parallel with the iris. In both conventional cases and Ex-Press cases, conjunctival suturing was performed with 10–0 nylon, and after confirming that there was no leakage from the bleb, the operation was terminated.
Transport and delivery of interferon-α through epithelial tight junctions via pH-responsive poly(methacrylic acid-grafted-ethylene glycol) nanoparticles
Published in Journal of Drug Targeting, 2019
Mary Caldorera-Moore, Julia E. Vela Ramirez, Nicholas A. Peppas
Transport studies were performed to determine the amount of IFN-α across a cellular monolayer with and without the presence of the P(MAA-g-EG-co-tBMA) nanoparticles. Passage 70 Caco-2 cells and passage 14 HT29–MTX cells were cultured in a 12-well Transwell® plates for 24 days as previously described. To prevent IFN-α from adhering to the Transwell® plates during the transport studies, the plates were first blocked with bovine serum albumin (BSA) (Sigma-Aldrich) prior to introducing IFN-α. A 0.5 mg·mL−1 BSA solution in Hank’s balanced salt solution (HBSS, Thermo Scientific, Waltham, MA) was used. The solution was then filtered using a GE Healthcare Whatman™ 0.02 µm anotop filter (Fisher Scientific, Waltham, MA) and pre-warmed for 20 min at 37 °C. Cell culture media was removed from the apical and basolateral chambers and replaced with pre-warmed BSA-HBSS solution after first washing both chambers one time with HBSS. The BSA-HBSS in both the apical and basolateral chambers contained Ca2+ at a concentration of 1.26 mM. Cells were allowed to equilibrate for 1 h and TEER measurements were taken at 0 and after 1 h. For all samples and TEER measurements the Transwell® plates were placed on a heating mat to maintain the temperature at 37 °C.