Precise and Facile Endotracheal Lung-tumor-implantation Mouse Model Visualized by GFP Expression.

BACKGROUND/AIM: In osteosarcoma, lung metastasis is a major cause of cancer-related death, as the 5-year survival rate for patients with metastases is approximately only 20-30%. To develop improved therapeutic strategies against lung-metastatic osteosarcoma, an experimental lung-tumor-implantation mouse model is needed for basic research. In the present study, we developed a precise and facile endotracheal lung-tumor-implantation technique. MATERIALS AND METHODS: For establishment of the lung-tumor-implantation mouse model of metastatic osteosarcoma, 5 mice were used. A 15-mm longitudinal incision was made in the center of the neck to expose the salivary glands. The salivary glands were then split, exposing the trachea covered by the sternohyoid muscles. The trachea was then clearly exposed by cutting the sternohyoid muscles longitudinally. A 22 G gavage needle was tilted slightly toward the left side of the mouse and inserted from the oral cavity into the bronchus, with confirmation of the position of the tip of the gavage needle visualized through the tracheal wall, followed by injection of 0.5% crystal violet to first confirm the accuracy of endotracheal injection in the lung. A 143B-GFP cell suspension (2.0×106 cells/50 µl PBS) was then injected endotracheally in other mice. RESULTS: The procedure, including anesthesia and suturing, took approximately 10 minutes. The left lobe of the lung, in which crystal violet was injected endotracheally, was stained in 3 out of 3 mice (100%). 143B-GFP-osteosarcoma tumors were detected with GFP fluorescence in the left lobe of the lung in 3 out of 4 mice (75%), 5 weeks after endotracheal injection. One mouse died 4 weeks after 143B-GFP-cell implantation. CONCLUSION: This novel technique of establishing tumors in the lung via endotracheal injection of cancer cells is precise and facile and can be used widely, since neither a surgical microscope nor X-ray imaging are needed.

In Vitro Evaluation of Cytotoxic Potential of Caladium lindenii Extracts on Human Hepatocarcinoma HepG2 and Normal HEK293T Cell Lines.

Data regarding the therapeutic potential of Caladium lindenii (C. lindenii) are insufficient. It becomes more important to explore plants as an alternative or palliative therapeutics in deadly diseases around the globe. The current study was planned to explore C. lindenii for its anticancer activity of ethanolic and hexane extracts of C. lindenii leaves against hepatic carcinoma (HepG2) and human embryonic kidney (HEK293T) cell lines. HepG2 and HEK293T cells were treated with 10, 50, 100, 200, and 400 µg/mL of ethanolic and hexane extracts of C. lindenii and were incubated for 72 h. Antiproliferative activity was measured by 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assay, and percentage viability were calculated through crystal violet staining and cellular morphology by Floid Cell Imaging Station. The study showed ethanolic extract exhibiting a significantly higher antiproliferative effect on HepG2 (IC50 = 31µg/mL) in a concentration-dependent manner, while HEK293T (IC50 = 241µg/mL) cells showed no toxicity. Hexane extract exhibited lower cytotoxicity (IC50 = 150µg/mL) on HepG2 cells with no effect on HEK293T (IC50 = 550µg/mL). On the other hand, the percentage viability of HepG2 cells was recorded as 78%, 67%, 50%, 37%, and 28% by ethanolic extracts, and 88%, 80%, 69%, 59%, and 50% by hexane extracts at tested concentrations of both extracts. Toxicity assay showed significantly safer ranges of percentage viabilities in normal cells (HEK293T), i.e., 95%, 90%, 88%, 76%, and 61% with ethanolic extract and 97%, 95%, 88%, 75%, and 62% with hexane extract. The assay validity revealed 100% viability in the control negative (dimethyl sulfoxide treated) and less than 45% in the control positive (cisplatin) on both HepG2 and HEK293T cells. Morphological studies showed alterations in HepG2 cells upon exposure to >50 µg/mL of ethanolic extracts and ≥400 µg/mL of hexane extracts. HEK293T on the other hand did not change its morphology against any of the extracts compared to the aggressive changes on the HepG2 cell line by both extracts and positive control (cisplatin). In conclusion, extracts of C. lindenii are proved to have significant potential for cytotoxicity-induced apoptosis in human cancer HepG2 cells and are less toxic to normal HEK293T cells. Hence C. lindenii extracts are proposed to be used against hepatocellular carcinoma (HCC) after further validations.

Naringenin suppresses epithelial ovarian cancer by inhibiting proliferation and modulating gut microbiota.

BACKGROUND: Ovarian cancer has the highest mortality among all gynecological malignancies; currently, no effective therapeutics are available for its treatment. Naringenin has been shown to inhibit the progression of various cancers, but its inhibitory effect on ovarian cancer remains unknown. PURPOSE: This study aimed to evaluate the inhibitory effects of naringenin on ovarian cancer and elucidate the underlying mechanisms. METHODS: Cancer cell proliferation was detected by cell counting kit-8 and crystal violet assays, and the migration capability was determined by wound healing and transwell assays. Western blotting and immunohistochemistry assays were employed to determine the expression levels of the epidermal growth factor receptor, phosphatidylinositol 3-kinase (PI3K) and cyclin D1 in vitro and in vivo, respectively. An ES-2 xenograft nude mouse model was established for the in vivo experiments, and fecal samples were collected for intestinal microbiota analysis by 16S rDNA sequencing. RESULTS: Naringenin suppressed the proliferation and migration of A2780 and ES-2 cancer cell lines and downregulated PI3K in vitro. In animal experiments, naringenin treatment significantly decreased the tumor weight and volume, and oral administration exhibited greater effects than intraperitoneal injection. Additionally, naringenin treatment ameliorated the population composition of the microbiota in animals with ovarian cancer and significantly increased the abundances of Alistipes and Lactobacillus. CONCLUSION: Naringenin suppresses epithelial ovarian cancer by inhibiting PI3K pathway expression and ameliorating the gut microbiota, and the oral route is more effective than parenteral administration.

Thoracoscopic Localization of Small Peripheral Pulmonary Lesions Using Percutaneous Computed Tomography-guided Pleural Dye Marking: A Retrospective Analysis.

Small pulmonary lesions are often difficult to localize during thoracoscopic surgery. We describe a new com-puted tomography (CT)-guided pleural dye-marking method for small peripheral pulmonary lesions that does not involve a visceral pleural puncture. We used this technique for 23 lesions (22 patients) who underwent tho-racoscopic partial lung resection (Nov. 2016-Jan. 2018). With the patient in the lateral decubitus position, pre-operative CT-guided marking on the skin over the lesion was performed. During the surgery, we marked the visceral pleura with a skin marker directly or with an infant-size nutrition catheter with crystal violet at the tip through a venous indwelling needle inserted perpendicular to the skin marking. We localized and resected the lesions in all cases, without complications. The median nodule size measured histopathologically was 8 (4-20) mm overall, and 7 (0-20) mm of the solid part; the median distance from the visceral pleura to the nodule was 9 (1-33) mm. The median operation time was 67 (37-180) min. The median postoperative hospital stay was 3 (3-11) days. Our CT-guided pleural dye-marking method is useful and safe for the localization of small periph-eral pulmonary lesions in thoracoscopic partial lung resections.

Review of the use of gentian violet in dermatology practice.

OBJECTIVE: To review the use of gentian violet in dermatology. DESIGN: A comprehensive literature search on gentian violet in dermatology practice was performed through PubMed. RESULTS: Gentian violet is effective in treating methicillin-resistant Staphylococcus aureus-colonized skin lesions; mean number of days for complete eradication was 9.1 days. Gentian violet is almost as effective as ketoconazole and more effective than nystatin in the treatment of oral thrush in AIDS patients. In an in vitro study on cutaneous T cell lymphoma cell lines, there was no difference between nitrogen mustard and gentian violet in stimulating apoptosis. When comparing gentian violet to silver sulfadiazine dressings in healing burn wounds, the gentian violet treatment group reported less pain, fewer febrile episodes, and decreased bacterial growth compared to control. In atopic dermatitis subjects, gentian violet decreased Staphylococcus aureus colonization and improved disease severity in lesional skin compared to non-lesional skin. CONCLUSION: Studies have investigated gentian violet's antibacterial, antifungal, antiviral, antiparasitic, anti-angiogenic, antitumor, and wound healing properties. Gentian violet is a low cost and well-tolerated topical agent with the potential for widespread applications in dermatology.

Regression of diffuse B-cell lymphoma of the leg with intralesional gentian violet.

In this case report, a patient of primary cutaneous diffuse B-cell lymphoma, leg type was treated with intralesional gentian violet as she was judged to be too medically fragile for conventional chemotherapy due to advanced age and multiple serious comorbidities. Gentian violet (crystal violet/hexamethyl pararosaniline) is a triphenylmethane dye. It has been shown to have an inhibitory effect on NADPH oxidase, an enzyme family which is found in abundance in reactive oxygen-driven tumors such as melanoma and lymphoma. We hypothesize that intralesional gentian violet treatment caused signalling changes in the lymphoma which allowed for immune clearance of the lymphoma. Complete resolution of the patient's lesion was noted on a follow-up visit.

In defence of 'dye therapy'.

Worldwide, healthcare is facing enormous problems with the continuing rise of drug-resistant infectious diseases. In view of the scarcity of new antimicrobial agents and the withdrawal of many pharmaceutical houses from the fray, alternative approaches are required. One of these is photoantimicrobial chemotherapy, which is highly effective across the range of microbial pathogens and does not suffer from resistance. However, there is a lack of uptake of this approach by healthcare providers and the pharmaceutical industry alike. It is seldom recalled that, unlike anticancer photodynamic therapy, the development of photoantimicrobial agents has evolved from the antiseptic 'dye therapy' in common use until the widespread introduction of the penicillin class in the mid-1940s. Cationic biological dyes such as methylene blue, crystal violet and acriflavine were effective in local wound therapy and today provide a sound basis for light-activated antimicrobial therapeutics. It is proposed that such 'safe' dyes are introduced as locally administered photoantimicrobials, especially in order to conserve valuable conventional antibacterial drugs.

Treatment strategies for aortic and peripheral prosthetic graft infection.

PURPOSE: The aim of this study was to investigate the outcomes correlated with our treatment strategy for prosthetic graft infection. METHODS: Seventeen patients were treated for prosthetic graft infections between 1997 and 2009. Initially, total graft excision was applied in five cases, partial graft excision was applied in six cases and graft preservation with drainage and irrigation was applied in six cases. Among the graft-preserved cases, four patients were infected with methicillin-resistant Staphylococcus aureus (MRSA) and treated with gentian violet (GV). RESULTS: The overall survival rate was 88% at 30 days and 82% at 1 year in this series. Of the excised cases, nine patients survived; however, two patients died. Among the cases in which MRSA-infected grafts were preserved, three patients survived; however, one patient died under a septicemic state. Infected graft preservation was applied at a high rate of 36%, and the mortality rate remained at 16%, without any signs of graft reinfection. CONCLUSIONS: In the treatment of infected grafts, the patient's condition should be considered in order to select the appropriate treatment in each case. Graft preservation should be considered as an alternative treatment option, especially in high-risk patients, and GV can be effective for conservative treatment of prosthetic graft infections, including MRSA infections.

Gentian violet: a 19th century drug re-emerges in the 21st century.

Gentian violet (GV) has a long and varied history as a medicinal agent. Historically used as an antibacterial and antifungal, recent reports have shown its utility as an antitypranosomal, antiviral and anti-angiogenic agent. The objective of this article is to summarize evidence regarding the efficacy and safety of GV use in dermatology. Recent discoveries have found novel targets of GV, namely NADPH oxidase in mammalian cells and thioredoxin reductase 2 in bacterial, fungal and parasitic cells. These discoveries have expanded the use of GV in the 21st century. Given that GV is well tolerated, effective and inexpensive, its use in dermatology is predicted to increase.

A simple and reproducible surgical technique for the management of preauricular sinuses.

Preauricular sinus is a relatively common congenital anomaly that mainly exists on the anterior aspect of the anterior limb of the ascending helix. Although many surgical techniques have been developed, extirpation of the sinus is not easy because of the ramifications of the sinus, remnants of the sinus wall, and infection with or without formation of abscesses, which can all lead to disease recurrence. In our institution, we have surgically treated a total of 141 cases of congenital preauricular sinuses. Instead of using the conventional lacrimal probe and methylene blue method, we used a gentian violet-soaked Cottonoid, which has antibacterial effects against the main pathogen responsible for causing infection of the preauricular sinus. Results have been very favorable, with a zero recurrence rate. We present here a simple and reproducible surgical technique using a gentian violet-soaked Cottonoid that even beginning surgeons can easily follow.

Major determinants of photoinduced cell death: Subcellular localization versus photosensitization efficiency.

We present a study on whether and to what extent subcellular localization may compete favorably with photosensitization efficiency with respect to the overall efficiency of photoinduced cell death. We have compared the efficiency with which two cationic photosensitizers, namely methylene blue (MB) and crystal violet (CV), induce the photoinduced death of human cervical adenocarcinoma (HeLa) cells. Whereas MB is well known to generate singlet oxygen and related triplet excited species with high quantum yields in a variety of biological and chemical environments (i.e., acting as a typical type II photosensitizer), the highly mitochondria-specific CV produces triplet species and singlet oxygen with low yields, acting mostly via the classical type I mechanism (e.g., via free radicals). The findings described here indicate that the presumably more phototoxic type II photosensitizer (MB) does not lead to higher degrees of cell death compared to the type I (CV) photosensitizer. In fact, CV kills cells with the same efficiency as MB, generating at least 10 times fewer photoinduced reactive species. Therefore, subcellular localization is indeed more important than photochemical reactivity in terms of overall cell killing, with mitochondrial localization representing a highly desirable property for the development of more specific/efficient photosensitizers for photodynamic therapy applications.

Disruption of the mitochondrial thioredoxin system as a cell death mechanism of cationic triphenylmethanes.

Alterations in mitochondrial structure and function are a hallmark of cancer cells compared to normal cells and thus targeting mitochondria has emerged as an novel approach to cancer therapy. The mitochondrial thioredoxin 2 (Trx2) system is critical for cell viability, but its role in cancer biology is not well understood. Recently some cationic triphenylmethanes such as brilliant green (BG) and gentian violet were shown to have antitumor and antiangiogenic activity with unknown mechanisms. Here we demonstrate that BG killed cells at nanomolar concentrations and targeted mitochondrial Trx2, which was oxidized and degraded. HeLa cells were more sensitive to BG than fibroblasts. In HeLa cells, Trx2 down-regulation by siRNA resulted in increased sensitivity to BG, whereas for fibroblasts, the same treatments had no effect. BG was observed to accumulate in mitochondria and cause a rapid and dramatic decrease in mitochondrial Trx2 protein. With a redox Western blot method, we found that treatment with BG caused oxidation of both Trx1 and Trx2, followed by release of cytochrome c and apoptosis-inducing factor from the mitochondria into the cytosol. Moreover, this treatment resulted in an elevation of the mRNA level of Lon protease, a protein quality control enzyme in the mitochondrial matrix, suggesting that the oxidized Trx2 may be degraded by Lon protease.

Da violeta de genciana à sorologia: o controle da transmissão da doença de chagas nos bancos de sangue / Gentian violet to serology: the control of transmission of Chagas disease in blood banks

Revisões históricas aos avanços científicos para o controle da doença, o Simpósio Internacional Comemorativo do Centenário da Descoberta da Doença de Chagas (1909-2009).

Salvador Mazza: considerado o redescobridor da doença de chagas pelos argentinos / Salvador Mazza: considered the chagas disease rediscoverer by Argentines

Revisões históricas aos avanços científicos para o controle da doença, o Simpósio Internacional Comemorativo do Centenário da Descoberta da Doença de Chagas (1909-2009).

RCT on gentian violet versus a hydrogel dressing for radiotherapy-induced moist skin desquamation.

OBJECTIVE: Radiotherapy-induced moist desquamation is a significant problem but there is a paucity of randomised data on which to base treatment decisions. The current prospective randomised trial compared gentian violet (GV) to a hydrogel dressing in this context. METHOD: Thirty patients undergoing radiotherapy to the head and neck region or breast who had developed moist desquamation in the radiotherapy field were randomised to treatment with 0.5% aqueous gentian violet (GV) (n=16) or a hydrogel dressing (n=14). The area of desquamation was regularly measured until healing or withdrawal from the study. RESULTS: The likelihood of healing with the hydrogel was greater than GV with a hazard ratio for healing of 7.95 (95% CI 2.20-28.68; p=0.002). The median time to healing for hydrogel was 12 days but had not been reached for GV by 30 days. Over the first 14 days the median'area under curve' of moist desquamation for GV was 82.6 cm2 (range 31.8-320.7 cm2) and that for hydrogel 20.0 cm2 (range 3.8-301.0 cm2) (difference significant at p=0.003). Ten of 16 patients treated with GV withdrew from the study (due to stinging in five and failure to heal in five) compared with two of the 14 treated with hydrogel (difference significant at p=0.021). CONCLUSION: Hydrogel dressings are more likely to heal radiotherapy-induced moist desquamation and are better tolerated than GV.

Targeted therapy of oral hairy leukoplakia with gentian violet.

Oral hairy leukoplakia (OHL) is a common oral manifestation of HIV infection. Clinically, these lesions appear as white plaques on the edges of the tongue. Pathophysiologically, these lesions occur because of infection of oral epithelium with Epstein-Barr virus (EBV). No universally effective therapy exists for OHL. We have previously shown that EBV infection and EBV viral products induce the generation of reactive oxygen. We have also demonstrated that the Food and Drug Administration-approved over-the-counter medication gentian violet is a potent inhibitor of reactive oxygen species. We thus chose to treat a patient with biopsy-proven OHL with topical gentian violet. Gentian violet solution was applied topically to the tongue of a patient with OHL. Complete clinical resolution was noted after three treatments. Treatment with topical gentian violet resulted in resolution of the lesions. Further studies with larger numbers of patients are required. The application of gentian violet can be used as a method to OHL treatment. Gentian violet is an inexpensive and safe therapy and, given that it inhibits reactive oxygen, this old therapy is now a targeted novel therapy.

Orthopaedic metal devices coated with a novel antiseptic dye for the prevention of bacterial infections.

Gendine is a novel antiseptic dye with broad-spectrum antimicrobial activity that may be used to coat plastics and metal devices. Our objective was to determine the efficacy of gendine-coated orthopaedic metal devices in preventing methicillin-resistant Staphylococcus aureus (MRSA) colonisation. Stainless steel and titanium Schanz rods were coated with gendine. The zone of inhibition (ZoI) around the rods with and without gamma-irradiation was determined by a modified Kirby-Bauer method. A previously published bioprosthetic biofilm colonisation model, modified Kuhn's method, was used to determine the adherence of MRSA to coated and uncoated rods, with and without irradiation, after insertion into bovine bone and after 3 months shelf life followed by 2 weeks of immersion in serum. The gendine-coated Schanz metal rods showed a net ZoI of 16 mm against MRSA before and after irradiation. Gendine-coated rods showed no biofilm formation (0 colony-forming units (CFU)), which was a significant reduction (P<0.001) compared with uncoated controls (>5000 CFU). Coated rods exposed to high-dose gamma-irradiation and coated rods drilled into bone also showed significant efficacy (P<0.001) in preventing biofilm adherence. After 2 weeks, gendine-coated rods maintained significant durability (P<0.01), resulting in 90% reduction in MRSA biofilm adherence compared with uncoated control rods. Results indicate that gendine-coated metal rods are highly efficacious in the prevention of MRSA biofilm.

Pleural space sterilization using gentian violet irrigation for postbullectomy empyema associated with artificial material infection.

Postoperative empyema associated with artificial material infection involves several treatment problems. We report the successful treatment of a case of post-bullectomy empyema with a small alveolar fistula that was associated with artificial material infection by Streptococcus viridans. In this case, complete empyema space sterilization was obtained by tube drainage and daily pleural irrigation using 0.1% gentian violet solution. This treatment circumvented the need for invasive surgery, including removal of the infected artificial materials and space-filling and/or collapse procedures. Consequently, gentian violet irrigation may be a useful treatment option in selected cases with complicated thoracic empyema.