Successful treatment of experimental murine vulvovaginal candidiasis with gentian violet.

Aim: This study evaluated the antifungal efficacy of gentian violet (GV) in an experimental vulvovaginal candidiasis (VVC) model. Materials & methods: In vitro susceptibility and cytotoxicity assays were performed to validate the antifungal potential and safety of GV. The antifungal efficacy was then evaluated in vivo through comparative analysis of the fungal burden following treatment with GV or nystatin, as well as assessment of the vaginal tissue by histology and electron microscopy. Results: GV demonstrated a safe antifungal profile against C. albicans, with a significant decrease in fungal burden and an improvement in the inflammatory process evaluated histologically. Conclusion: The results of this study motivate further assessment of GV as a promising alternative for VVC therapy.

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.

Lugol's solution and Gentian violet eradicate methicillin-resistant Staphylococcus aureus biofilm in skin wound infections.

The study aimed to evaluate the antibacterial efficacy of Lugol's solution 5% and Gentian violet 1% against methicillin-resistant Staphylococcus aureus (MRSA) biofilm in vivo. The bactericidal efficacy for treatment of MRSA-biofilm skin wound infection was tested in a murine model. Luciferase-tagged S. aureus Xen31, a MRSA-strain derived from S. aureus ATCC-3359130, was used for infection. Wounds were made in the skin of mice and infected with MRSA. The mice were treated with Lugol's solution and Gentian violet. Application of the antimicrobial agents started 24 hours post infection and was repeated daily for five-days. The antimicrobial effect on the biofilm bacteria was evaluated by measuring bioluminescence from MRSA daily for seven-days. Lugol's solution and Gentian violet showed a significant reduction in luminescent signals from the first assessment day to all subsequent days (P < .001). Lugol's solution and Gentian violet effectively eradicated MRSA in biofilm in vivo and could be alternatives or in addition to topical antibiotics when MRSA-biofilm wound infection is suspected.

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.

Effect of intravaginal gentian violet for acute vaginal candidiasis treated with a single dose oral fluconazole: a randomised controlled trial.

The study aims to evaluate the additive effect of intra-vaginal gentian violet (GV) on a single dose oral 200 mg fluconazole for acute vaginal candidiasis (VC). Women aged ≥18 years who had VC were randomly allocated to receive either fluconazole 200 mg (group 1, FLU, N = 90); or the fluconazole with GV (group 2, FLU + GV, N = 93). Outcome measures were 2-week clinical cure rate, conversion of positive fungal culture, time-to-cure, side effects, satisfaction and symptomatic recurrence within 2 months. No significant difference of participants' characteristics was observed. They were 32.4 ± 8.7 year-old and non-obese. Participants receiving FLU + GV had higher clinical cure rates (81.7% vs. 74.4%, p=.236); lower recurrence rate (19.4% vs. 30.0%, p=.097); shorter time-to-cure (3.1 vs. 4.0 days, p=.013); but lower culture conversion rate (74.2% vs. 80.0%, p=.351). Participants in both groups reported high satisfaction and none had severe adverse events. In conclusion, the addition of GV results in a shorter time-to-cure but not cure rate. Clinical trial registration: TCTR20180917003 (http://thaiclinicaltrials.org/show/TCTR20180917003).Impact StatementWhat is already known on this subject? The efficacy of fluconazole for acute vaginal candidiasis is limited to 75-90% due to drug resistance and non-albicans Candida. Gentian violet (GV) has long been used for mucosal candidiasis; and is recommended as the second line treatment for women with recurrent vulvovaginal candidiasis (RVVC).What do the results of this study add? Adding GV to a single oral 200 mg fluconazole results in a quicker resolution of symptoms of acute VC but not cure rate. The participants' satisfaction and acceptance are high. Lifestyle modification, particularly reduction of sugar-rich diet, associates with the higher culture-based cure rate.What are the implications of these findings for clinical practice and/or further research? As GV is widely and easily accessible, and speculum examination with or without microscopy is the main diagnostic tool of VC; the single application of GV seems doable in real-life practice. This simple anti-septic solution can accelerate symptom resolution. However, the proper frequency of GV application should be further explored. As importantly, lifestyle modification should always be included in counselling session to optimise treatment outcome.

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.

Management of a Complex Peristomal Calciphylaxis: A Case Study.

BACKGROUND: Calciphylaxis, also referred to as calcific uremic arteriolopathy, is a rare and serious syndrome of small blood vessels with a high mortality rate. Calciphylaxis lesions require intensive wound management with medical interventions for the patient to survive the sequelae of sepsis and present unique challenges when found in the peristomal skin. CASE: A 33-year-old man presented with multiple malodorous stage 4 pressure injuries of his sacrococcygeal and gluteal area. His medical history included chronic kidney disease requiring hemodialysis since 2007, diabetes mellitus, and incomplete paraplegia. He underwent diverting colostomy to enhance wound healing. His hospital stay was complicated by the development of a peristomal calciphylaxis lesion (PCL) that made ostomy pouching especially challenging for the nursing staff. His care needs were also aggravated by nonadherence to diet restriction, pressure injury prevention efforts, and a continued need for high doses of analgesic medication. Collectively, these issues presented a challenge for the health care team during his hospital course and during safe discharge planning. CONCLUSIONS: The peristomal calciphylaxis lesion decreased in surface area and improved in appearance with the use of various wound care products as his medical condition improved. Skilled nursing management in the context of ongoing interdisciplinary collaboration assisted in managing the patient's peristomal calciphylaxis, ultimately leading to safe discharge from hospital.

A prospective evaluation of methylene blue and gentian violet dressing for management of chronic wounds with local infection.

The objective of this prospective, non-randomised study was to evaluate the performance of an antibacterial foam dressing containing methylene blue and gentian violet (Hydrofera Blue Classic dressing® ) for the management of chronic wounds with local infection. Patients in this study were ≥18 years of age (n = 29), and each had at least one chronic wound ≥1 cm2 in size that showed signs of localised infection or critical colonisation but with good potential for healing based on clinical assessment. To all of these wounds, the dressing was applied and changed three times per week over the 4-week study period. The primary endpoints of the study were: (i) changes in wound surface area measurement, (ii) changes in Pressure Ulcer Scale for Healing (PUSH) scores, (iii) changes in percent surface area of devitalised tissue (i.e., yellow slough or other necrotic tissue) and (iv) changes in clinical signs associated with localised wound infection/critical colonisation. Participants were evaluated at presentation (week 0 = baseline), week 2 and at week 4 (end of the study). The 29 patients completed the study, and at week 4, the following wound improvements were observed: (i) baseline mean wound surface area was significantly reduced by 42·5%, from 21·4 to 12·3 cm2 at week 4 (P = 0·005); (ii) baseline mean PUSH score decreased significantly from 13·3 to 10·7 at week 4 (P < 0·001); (iii) baseline mean wound coverage by devitalised tissue (%) was significantly reduced, from 52·6 % to 11·4% at week 4 (P < 0·001) and (iv) the mean UPPER and LOWER wound infection scores were reduced from 3·6 at baseline to 0·9 at week 4 (75%; P < 0.001). These results indicate that the Hydrofera Blue Classic dressing was effective at managing these chronic wounds and helped them progress onto a healing trajectory.

Use of Ovine-based Collagen Extracellular Matrix and Gentian Violet/Methylene Blue Antibacterial Foam Dressings to Help Improve Clinical Outcomes in Lower Extremity Wounds: A Retrospective Cohort Study.

Dressings that provide broad spectrum metalloprotease reduction along with inherent aspects of an extracellular matrix may contribute to improved wound healing outcomes and shorter treatment times. OBJECTIVE: The author performed a retrospective case series analysis to determine the clinical outcomes of regular debridement with the use of ovine-based collagen extracellular matrix dressings and gentian violet/methylene blue polyurethane antibacterial foam dressings in treating 53 patients with 53 chronic lower extremity wounds (diabetic foot ulcers [DFUs], venous leg ulcers, and heel pressure ulcers). MATERIALS AND METHODS: Patients were treated twice weekly in an outpatient clinic for the first 4 weeks and weekly thereafter until closure. RESULTS: Average body mass index (BMI) for the study population was 28.3, and the average patient age was 75.9 years. Mean percent wound surface area reduction at 4, 8, and 12 weeks was 38.5%, 73.3%, and 91.3%, respectively. Average time to closure for all wounds was 10.6 weeks (range, 5-24 weeks). All wounds were 100% reepithelialized by week 20 except 1 DFU that reepithelialized at week 24. The average cost of care for a single wound episode (from presentation to closure) was $2749.49. CONCLUSION: Results of this analysis showed that the healing of chronic wounds in this series could be achieved at a reasonable cost with regular debridement and a collagen matrix dressing regimen, even in patients of advanced age and above average BMI as well as in wounds that did not achieve > 40% wound surface area reduction at 4 weeks.

Use of antiseptic for cord care and its association with neonatal mortality in a population-based assessment in Bihar State, India.

OBJECTIVES: Dry cord care is recommended for all births by the Health Ministry in India. We report prevalence of antiseptic cord care in the context of neonatal mortality in the Indian state of Bihar. DESIGN: Population-based cross-sectional study with multistage stratified random sampling. SETTING: Households in 1017 clusters in Bihar. PARTICIPANTS: A representative sample of 12 015 women with a live birth in the last 12 months were interviewed from all 38 districts of Bihar (90.7% participation) in 2014. PRIMARY AND SECONDARY OUTCOME MEASURES: Use of antiseptic cord care at birth and its association with neonatal mortality using multiple logistic regression. RESULTS: Topical application of any material on cord was reported by 6534 women (54.4%; 95% CI 53.5% to 55.3%). Antiseptic cord care prevalence was 49.7% (95% CI 48.8% to 50.6%), the majority of which was gentian violet (76.4%). The odds of antiseptic use for cord care were higher in facility births (OR 1.46; 95% CI 1.27 to 1.69) and for deliveries by a qualified health provider (OR 1.44; 95% CI 1.26 to 1.66), but were lower for births that occurred before the expected delivery date (OR 0.77; 95% CI 0.61 to 0.96). A total of 256 (2.1%) newborns died during the neonatal period. The odds of neonatal death were significantly higher for live births with no reported antiseptic use (OR 1.53; 95% CI 1.18 to 1.99), and this association persisted when live births in health facilities were considered separately. CONCLUSIONS: Even though dry cord care is recommended by health authorities in India, half the women in this study reported use of antiseptic for cord care mainly with gentian violet; and its use had beneficial effect on neonatal mortality. These findings suggest that the application of readily available gentian violet for cord care in less developed settings should be assessed further for its potential beneficial influence on neonatal mortality.

Treatment of extensive erythema multiforme with topical gentian violet.

Topical and systemic therapies for erythema multiforme have been widely described in the literature. The pathogenesis of erythema multiforme involves increased expression of vascular endothelial growth factor resulting in the promotion of microvascular permeability and angiogenesis. Gentian violet has been shown to have antiangiogenic properties. Here, we present a case of erythema multiforme successfully treated with topical gentian violet. We report the case of a patient who presented with erythema multiforme. Prior pertinent history included diabetes mellitus type I, limiting the clinical use of systemic corticosteroids. Topical gentian violet was used to treat the cutaneous lesions. Our patient responded well to treatment with topical gentian violet with stabilization and resolution of the lesions without using systemic therapy that may pose serious side effects in the setting of other comorbidities such as diabetes mellitus type I. This case highlights the variable therapeutic options available for treatment of erythema multiforme, including topical gentian violet. While further studies are needed, this case demonstrates the antiangiogenic properties and clinical utility of topical gentian violet in the treatment of erythema multiforme.

Gentian Violet and Methylene Blue Polyvinyl Alcohol Foam Antibacterial Dressing as a Viable Form of Autolytic Debridement in the Wound Bed.

The objective of this article is to describe the results of a comparative porcine study that evaluated the effectiveness of a gentian violet and methylene blue (GV/MB) polyvinyl alcohol (PVA) antibacterial foam dressing in debriding eschar. The authors performed an in vivo, preclinical study on eschar-covered porcine wounds. Two clinical case studies are also included. Test products, GV/MB antibacterial foam dressing, collagenase ointment, collagenase ointment plus GV/MB antibacterial foam dressing, medical-grade honey, and moist gauze dressing (control), were applied to porcine wounds using a split-back study design. The percent of eschar removal and wound closure were measured and recorded at time points up to 14 days. Statistically significant reduction in eschar was observed with GV/MB dressing and with GV/MB dressing with collagenase. By day 14, the wounds with GV/MB dressing alone and GV/MB dressing with collagenase had eschar covering less than 25% of the wound bed area compared with collagenase alone, medical grade honey, or moist gauze control, which showed eschar still covering over 75% of the wound bed area. Autolytic debridement activity of GV/MB foam dressings was evident in the porcine eschar study, as well as in the cases described.

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.