Methylene Blue Staining Method Research Articles

Surface modification of polyester fabrics using choline hydroxide-catalysed glycolysis

This study explores the surface modification of polyethene terephthalate (PET) fabric via glycolysis, employing choline hydroxide as an eco-friendly catalyst. A range of chemicals was evaluated for catalysing the glycolysis reaction. Among them, choline hydroxide demonstrated notable enhancements in surface hydrophilicity, moisture regain, wicking properties, and water contact angle, surpassing other catalysts. Scanning Electron Microscopy (SEM) revealed a marked increase in surface roughness and irregularities following glycolysis in the presence of this ionic liquid. The surface modification technique employed in this investigation resulted in a decrease of less than 20% in the breaking force of the fabric. Fourier Transform Infrared (FT-IR) spectroscopy and the methylene blue staining method confirmed the emergence of free hydroxyl groups on the modified fabric's surface. These findings suggest that glycolysis catalysed by choline hydroxide is a promising approach for enhancing the surface characteristics of polyester fabrics, which may lead to improved functional properties in textile applications.

Comparison of bronchial methylene blue staining and modified inflation-deflation method in identifying the intersegmental plane during lung segmentectomy.

Identification of the intersegmental plane (ISP) is the critical step in lung segmentectomy because of the complicated anatomic variations. Bronchial methylene blue staining was developed by our team in 2015 and is now commonly used at our center, it could rapidly and accurately identify the ISP. In this study, we aimed to compare bronchial methylene blue staining with the modified inflation-deflation method in terms of their perioperative characteristics and to present our experience of the methylene blue method. From June 2020 to September 2021, the data of 112 patients with pulmonary ground-glass nodules who underwent segmentectomy by video-assisted thoracoscopic surgery were retrospectively reviewed. Sixty-two patients underwent bronchial methylene blue staining, and 50 patients underwent the modified inflation-deflation method. Both methods could accurately identify the ISP. The time taken to clearly display the ISP (82.94±28.08 vs. 868.20±145.89 seconds; P<0.001) and the surgical duration (131.69±32.05 vs. 146.08±28.11 minutes; P=0.014) were significantly shorter in the bronchial methylene blue staining group than in the modified inflation-deflation group. There were no significant differences between the two groups in the bleeding volume, drainage time, and length of postoperative hospital stay, as well as in most other perioperative characteristics. Compared with the modified inflation-deflation method, the bronchial methylene blue staining method can quickly display the ISP and shorten the surgical duration. This method is safe and feasible, can be widely applied during thoracoscopic anatomic segmentectomy.

Preoperative localization of sentinel lymph nodes using percutaneous contrast-enhanced ultrasonography in patients with breast cancer.

This study aimed to investigate the feasibility of preoperative identification of sentinel lymph nodes (SLNs) by contrast-enhanced ultrasound (CEUS) for patients with breast cancer. The patients with T1-T2N0M0 breast cancer who were scheduled for primary surgical treatment were recruited. All the patients had received a periareolar intradermal injection of an ultrasonic contrast agent (SonoVue, Bracco, Milan, Italy) followed by an ultrasound to identify contrast-enhanced SLNs. A guidewire was deployed to localize the SLN. Methylene blue stain was used to help trace SLNs during the operation. The identification rate and accuracy rate were recorded. The number of SLNs labeled by two methods was counted and compared using Wilcoxon testing. A total of 366 SLNs were detected in 72 patients by methylene blue intraoperatively, with a median of 5 lymph nodes [interquartile range (IQR), 4-6] per patient. A total of 95 SLNs were detected in 63 patients (87.5%) by CEUS, with a median of 1 lymph node (IQR, 1-2) per patient. The number of SLNs detected by CEUS was significantly less than that labeled by the methylene blue staining method (Z=-7.362, P=0000). Pathology confirmed 12 single metastases in all the lymph nodes examined, 10 of which were the only lymph node identified by CEUS. Periareolar intradermal injection of an ultrasonic contrast agent was an effective and convenient supplementary to localize SLNs. The technique was expected to improve the accuracy of axillary staging with minor surgical trauma and postoperative complications.

Diagnostic efficiency of methylene blue and lugol's iodine double staining method in oral leukoplakia in detecting dysplasia.

Early detection of dysplasia in oral potentially malignant disorders (PMD) might facilitate screening for possible subsequent malignant transformation. Vital staining is a non-invasive clinical adjunct used for determining the biopsy site, which facilitates early detection of dysplastic changes in PMD. Some authors suggested that double staining method has superior results over staining with a single dye. The aim of the study was to evaluate the accuracy of in vivo staining with methylene blue (MB) and Lugol's iodine (LI) double staining method in comparison with MB staining alone. Fifty patients of oral leukoplakia were recruited for the study. After obtaining written informed consent from the patients, the lesions were stained consecutively with 5% MB and 3% LI. The pattern of dye retention of MB alone, followed by MB and LI was noted. Incisional biopsy from the lesion was taken based on the retention of MB and the absence of staining of LI or by clinical judgement in case both stains were not retained. The clinical uptake of the stains was correlated with the degree of dysplasia on histopathological examination. Out of 50 subjects, MB was retained in 47 cases (94%), while 3 cases (6%) failed to retain the dye. However, out of 47 cases, 20 cases had dark blue stain and were considered as MB positive, while the rest 27 cases had pale blue stain and were considered to be negative for MB staining. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic accuracy (DA) of MB staining were 57.82%, 100%, 100%, 26.53% and 46%, respectively. After applying both stains, i.e., double staining method, the outline of the lesion was better defined. Out of 50 PMD cases, 3 patients retained only LI and showed no dysplasia. Out of 47 cases which showed dysplasia, 27 cases showed pale blue retention of MB (-) and also were negative for LI. The rest 20 cases were positive for MB but negative for LI. The sensitivity, specificity, PPV and NPV with double staining technique were 100%. The accuracy and reliability of double staining method was superior to MB staining. Increased accuracy of double staining method aids in better detection of dysplasia and is of great help to the clinician in deciding the nature of PMD in question.

Optimization of in vitro pollen germination and pollen viability tests for Castanea mollissima and Castanea henryi

The Castanea Miller (Fagaceae) family are commercially important trees in the word, grown for their nuts that are rich in starch and nutrients. The production of nuts is related to pollen fertility, but little is known about pollen viability and germination of the family. In this paper, we investigated the staining and in vitro germination traits of Chestunt (Castanea mollissima Blume) and Chinese Chinquapin (Castanea henryi (Skam) Rehder & Wilson), which were widely cultivated in China. We aimed to determine the optimal media for pollen germination and to develop a method to estimate viability using a series of correlation analyses based on staining and germination results. Pollen viability was assessed using 2,3,5-triphenyl tetrazolium chloride (TTC), 2,5-diphenyl monotetrazolium bromide (MTT), acetic carmine, benzidine-H2O2, blue ink dyeing, methylene blue, I2-KI, inorganic acid, and peroxide staining methods. For germination tests, different concentrations of sucrose, boric acid (H3BO3), calcium chloride (CaCl2), and gibberellin (GA3) were used. Benzidine-H2O2 and MTT dyes were determined to be suitable for the pollen viability test. The optimal media compositions for C. mollissima and C. henryi both had 5% sucrose, 0.01% H3BO3, and 0.005% GA3, but their amounts of CaCl2 varied at 0.01 and 0.005%, respectively. Assessments of staining methods found that MTT staining exhibited a linear relationship with pollen germination (***p < 0.001), which overestimated viability by an average of 1.4× for C. mollissima and an average of 1.7× for C. henryi, compared to in vitro germination rates. The proportion of dark staining by MTT was close to the germination rate. This study has identified optimal media and a method to estimate germination viability, which will improve Castanea Miller breeding programs.

Application of the Reactive Oxygen-Based Cervical Exfoliated Cell Staining Solution in Clinical Cervical Cancer Screening

We developed a special methylene blue solution for staining of cervix shedding cells based on catalytic oxidizing chromogenic reaction, which shows a potential for cervical cancer cytology screening. We screened a total of 1 922 women for cervical cancer with the special methylene blue staining method and a conventional Pap smear method using cervix shedding cells. Then, the patients with positive indicators of the Pap smear or this special solution staining method were examined by the electron colposcopy and histopathological examination. Staining of cervical exfoliated cells with this reactive oxygen-based special solution showed that the number of positive cases was 140 (7.28%). Among them, 21 cases showed dark green (1.09%), and 119 cases showed purple black (6.19%). The results of the Pap smear method showed that the number of positive cases was 123 (6.40%), of which ASCUS was 105 (5.46%), ASC-H was 5 (0.26%), and LSIL was 9 (0.47%), and HSIL was 4 (0.21%). For cervical exfoliated cell special staining solution for screening cervical intraepithelial neoplasia (CIN-?, CIN-?) and cervical cancer, sensitivity was 83.33%, specificity was 65.52%, accuracy was 74.29%, missed diagnosis rate was 13.33%, positive coincidence rate was 51.43%, and the negative coincidence rate is 86.67%. Our results proved the value of this method for early screening of cervical cancer through clinical practice in China.

Preliminary studies on the anti-osteoporosis activity of Baohuoside I

The aim of the present study was to investigate the anti-osteoporotic activity of Baohuoside I, an active component of Herba Epimedii. Effects of Baohuoside I on the differentiation of BMSCs and the formation of adipocytes were evaluated using alkaline phosphatase staining and methylene blue staining method, respectively. Osteoporosis model was established in ovariectomized rats prior to the measurement of the serum SOD and MDA levels as well as the expression of inflammatory cytokines protein in the rats’ tissues after treatment with Baohuoside I using ELISA assay kits. The estrogen-like effect of Baohuoside I was also measured on HeLa cells. The positive rates of ALP staining in Baohuoside I groups were significantly higher (p < 0.01) compared with the normal group, with no obvious adipocyte formation observed in the groups that received Baohuoside I treatments. The levels of inflammatory markers (IL-1β, TNF-α, IL-6 and IL-8) in the treated groups were significantly lower (p < 0.05) than in the model group. Likewise, the treated groups exhibited a significantly higher (p < 0.05) serum levels of MDA compared with the model group, while SOD levels were markedly lower (p < 0.05) in a dose-dependent fashion. Baohuoside I showed no estrogen-like effect on HeLa cells upon treatment with the drug. Collectively, these results indicated that the anti-osteoporotic activity of Baohuoside I could be related to the induction of BMSCs differentiation into osteoblasts coupled with the inhibition of adipocyte formation, regulation of immune functions, and antioxidant activity.

Better detection of Demodex mites by Löffler's alkaline methylene blue staining in patients with blepharitis.

PurposeTo determine whether the Löffler’s alkaline methylene blue staining method is better than no staining in detecting Demodex mites in the eyelashes of patients with blepharitis.Materials and methodsEyelashes were collected from 22 patients with blepharitis. The mean age of the patients was 82.5±6.2 years (± SD) with a range from 71 to 93 years. Eyelashes were epilated by forceps and placed individually on microscope slides. The number of Demodex mites was determined by conventional optical microscopy before and immediately after the addition of the methylene blue staining solution.ResultsThe mean Demodex count before the addition of the methylene blue solution was 2.9±2.9, and it was 4.4±3.9 after the addition of the methylene blue solution (P<0.01, Wilcoxon test).ConclusionThe methylene blue staining method is a simple and useful method in detecting the presence and quantifying the number of Demodex mites. We recommend the methylene blue staining method not only for the diagnosis of the presence of Demodex mites but also to evaluate the therapeutic effects of medications to eliminate the mite infestation.

Methylene Blue Staining Test in Early Detection of Head and Neck Cancers

Head and neck cancers represent an important health problem in the present time. Due to the often late diagnosis and the significant changes that accompany the therapy associated with this disease, the impact on the quality of life of the patient is remarkable, causing a lower adherence to treatment, poor outcomes and an unfavorable prognosis. The current trend focuses on methods of investigation that allow an early diagnosis, which in turn translates into minimally invasive interventions with a favorable outcome. The purpose of this paper is to present the methylene blue staining method as an diagnosis tool, allowing the surgeon a better evaluation of the tumor and a more reliable limit of resection.

Factor analysis and method exploring for lymph nodes harvest in gastric cancer

The number of lymph node dissection and positive lymph nodes are the necessary guarantees for patients to achieve accurate staging after gastric cancer surgery. On the basis of the minimum number of lymph nodes dissection recommended by the NCCN guidelines, as many as possible lymph node yields will be most likely to benefit patients. Many factors can influence the number of lymph node yields including surgery, patient, tumor pathology and postoperative sorting factors. Compared with traditional manual nodal dissection method, fat-clearing technique and methylene blue staining method can improve the number of lymph nodes detection, while lymphatic tracers, such as carbon nanoparticles, are conducive to show lymphatic vessels, contributing to the dissection of small lymph nodes. The initial results from People's Liberation Army General Hospital show that lymph node packet submission after isolation by surgeon yields more lymph nodes. For the establishment of standards, lymph node retrieval-related procedures need further in-depth exploration and investigation.

넙치에 기생하는 쿠도아 점액포자충(Kudoa septempunctata) 의 활성 확인을 위한 염색법의 비교

Consumption of Paralichthys olivaceus in raw fish have been reported as the cause of outbreaks of food-born illness, and Kudoa septempunctata in muscle of Paralichtys olivaceus was suggested with the causative agent. For this reason, distinguish of vital and dead spore is important to study survivability of Kudoa septempunctata in human intestinal condition and in vitro inactivation of Kudoa septempunctata. In this reports, we suggest NR & MB (Neutral red and Methylene blue) staining method that is easier and simpler than the previously described HO & PI (Hoechst33342 and Propidium iodide) method according to a experimental condition.

Rapid alkaline methylene blue supravital staining for assessment of anterior segment infections.

PurposeTo present the Löffler’s alkaline methylene blue technique of staining eye discharges in eyes with anterior segment infections.MethodThe Löffler’s alkaline methylene blue staining method is a simple staining technique that can be used to differentiate bacterial, viral, and fungal infections. It is a cationic dye that stains cells blue because the positively charged dye is attracted to negatively charged particles such as polyphosphates, DNAs, and RNAs. Specimens collected from patients by swabbing are smeared onto microscope slides and the methylene blue solution is dropped on the slide. The slide is covered with a glass cover slip and examined under a microscope. The entire time from the collection to the viewing is about 30 seconds.ResultsHistopathological images of the conjunctival epithelial cells and neutrophils in eye discharges were dyed blue and the nuclei were stained more intensely blue. Bacterial infections consisted mainly of neutrophils, and viral infections consisted mainly of lymphocytes.ConclusionsLöffler’s alkaline methylene blue staining can be done in about 30 seconds for diagnosis. Even though this is a one color stain, it is possible to infer the cause of the infection by detection of the absence of bacteria and/or fungi in context of the differential distribution of neutrophils and lymphocytes.

Methylene Blue Staining: A New Technique for Identifying Intersegmental Planes in Anatomic Segmentectomy

Pulmonary segmentectomy is being increasingly used to resect small lung nodules; however, identifying the intersegmental plane is difficult. We describe a new methylene blue staining technique that we developed to identify the intersegmental planes in anatomic segmentectomy using video-assisted thoracic surgery (VATS) or thoracotomy and to evaluate its feasibility and safety. Between October 2013 and December 2013, 14 consecutive patients with lung disease underwent anatomic segmentectomy at our institution (10 VATS, 4 conventional thoracotomy). Methylene blue 0.1% (20 mL) was slowly injected into the bronchus of the target pulmonary segments using an intravenous needle after division of the artery, vein, and bronchus of the target segments, and the boundaries were detected, followed by anatomic segmentectomy. The staining took only 3 min. The target pulmonary segments stained blue, allowing for the clear identification of the intersegmental plane on both the surface and in the lung parenchyma, and all operations were successfully completed. Staining did not affect pathologic examination of the resected specimens. The fluid that drained from the chest tube and the patients' sputum, urine, and feces were not blue. There were no perioperative deaths or major complications. To our knowledge, this study is the first to report a safe and feasible methylene blue staining method for identifying the lung segment borders that does not require any special equipment. More importantly, this method can clearly detect the intersegmental planes on the pleural surface and within the lung parenchyma, enabling thoracic surgeons to accurately perform anatomic segmentectomy.

The value of simple microbiological studies for on-site screening of acute neonatal conjunctivitis in Angola

BackgroundNeonatal conjunctivitis or ophthalmia neonatorum (ON) is an acute bacterial conjunctivitis contracted by newborns during delivery. In non-industrialized countries, detection of the etiological agent is difficult due to the unavailability of modern diagnostic resources. Therefore, we analyzed the effectiveness of Gram and methylene blue staining techniques, which are simple microbiological methods in suspecting the aetiology of ON in a maternity ward in Luanda, Angola.FindingsNeonatal conjunctival smears (n = 95), maternal data, and perinatal factors were collected. Slides were air-dried and sent to the Microbiology Department of the Hospital Clinico Universitario, Valladolid, Spain, where trained personnel performed Gram and methylene blue staining methods. Findings were interpreted by two expert microbiologists. Ophthalmological examination of all children showed five newborns with clinical signs of ON. Fourteen mothers reported were suspected with vulvo-vaginitis, and 27 had a urinary infection during pregnancy. Gram staining revealed the presence of epithelial cells in 87.6% and leukocytes in 15% of the conjunctival smears. These values were significantly higher than those shown by methylene blue staining. No rods, cocci, or yeasts were identified by either staining method. Chlamydia trachomatis DNA was also undetected in a small sub-sample with clinical suspicion of ON. There was no correlation among the presence of ON, ON microbes, maternal data, or perinatal factors.ConclusionsBasic microbiological techniques did not provide enough information for screening cases of ON in Angola. Therefore, the use of molecular biology or other techniques is warranted for this purpose.

A novel antimicrobial photosensitizer: hydroxyethyl Michler’s ketone

A new photoactive antimicrobial agent was synthesized by introducing a reactive functional group (–CH2CH2OH) to 4,4′-bis(dimethylamino)benzophenone (Michler’s ketone) that could enhance the fixation of benzophenone derivatives to cotton fabric. Cotton fabrics containing organic photosensitizers were investigated to demonstrate antimicrobial properties in the application to protective and germ-free clothing. Photosensitizers such as benzophenone, Michler’s ketone and synthesized (4-(dimethylamino)phenyl)(4-((2-hydroxyethyl)(methyl)amino)phenyl)methanone (MK-EtOH) were applied on cotton fabrics through a pad–dry–cure method. The introduction of the reactive functional groups (–CH2CH2OH) enhanced the fixation of benzophenone derivatives to cotton fabrics as confirmed by Fourier transform infrared spectroscopy analysis and methylene blue staining method. No significant reductions of tensile strength and thermal degradation were observed with cotton fabrics treated with MK-EtOH. The scanning electron microscope morphology of the treated cotton showed that organic photosensitizers were evenly distributed and diffused to cotton fibers. After ultraviolet irradiation (365 nm), the cotton fabrics treated with synthesized MK-EtOH photosensitizer provided more effective and durable antimicrobial properties than Michler’s ketone and benzophenone, even after multiple washing.

English

This study is on&nbsp;the multipotential mesenchymal cells which enable us to differentiate chondroblasts in presence of&nbsp;Transforming growth factor&nbsp;(TGF&beta;) andbone morphogenic factors&nbsp;(BMPs). As use of a transferring system for these growth factors is a notable issue in recent researches, the present study was designed to evaluate the nanosilver transferring system of TGF&beta; on two sample groups including 42 rats and 4 rabbits as pilot study. The samples were divided in two groups for injection of TGF&beta; with and without nanosilver. Femoral muscle of rats were injected and then sacrificed after 1, 2 and 4 weeks. This injection was repeated in subperiosteal area of parietal bone of rabbits with decreased dose of nanosilver. The injected areas were examined histopathlogically using H&amp;E and Methylene-blue staining methods. Results revealed chondrogenesis was not detected in all 6 rat groups in all following weeks, but was present in the parietal bone of one rabbit which was injected with TGF&beta; with nanosilver and sacrificed after 4 weeks. The differences in level of inflammation in and presence of necrosis were significant in TGF&beta; with nanosilver groups vs TGF ones. Although, addition of nanosilver to TGF&beta; did not show great cytotoxic responses, it may not have great effects to induction of chondrogenesis in rat femoral muscle. Detection of chondrogenesis in one sample of subperiosteal rabbit parietal bone can be explained by decreased nanosilver dose and difference in anatomic site of injection. So, more investigations need to verify practical use of TGF&beta; with nanosilver transferring system in repair of chondral defects. &nbsp; Key words:&nbsp;Transforming growth factor (TGF&beta;), bone morphogenic factor (BMP2), nanosilver, chondrogenesis.

Quality Improvement of the Human Hair by the Treatment of Protease Extracted from Earthworm

The feasibility of earthworm protease as a new cosmetic material for human hair care was investigated. The effectiveness of the earthworm protease treatment was assessed by thickness change of hairs, optical microscope examination, aminoacid analysis, surface morphology, angular resolution analysis through methylene blue staining method and tensile strength change. The protease treated hair became thinner and the soil on the surface removed in experimental groups unlike control group. Tensile strength decreased in experimental group in which the enzymes may decompose polypeptide bonds.

Assessment of yeast viability under hyperbaric conditions through a modeling approach

AbstractThe effect of pressure (0.1–1.5 MPa) and oxygen concentration on the growth and viability of Saccharomyces cerevisiae was studied. Cell viability was assessed through the Methylene Blue staining method and the percentages of viable and non‐viable cells were estimated using digital image processing. A model taking into account cell viability was developed and used to describe the measured data. The model reveals the opposing effects between oxygen availability and the baric and oxidative stresses present on the system and can successfully describe not only the traditional biomass–product–substrate (X–P–S) evolution but also the ratio of viable cells with time. It is shown that cell viability in general is not constant during the experiments but strongly depends on the environment. Copyright © 2005 Society of Chemical Industry

Synergistic antitumor activity of troxacitabine and camptothecin in selected human cancer cell lines.

Troxacitabine (L-OddC) is an L-configuration deoxycytidine analog currently in phase II trials for the treatment of cancer. The cytotoxicity of L-OddC in combination with other anticancer agents has not been studied systematically. In the present study, we assessed the cytotoxic effects produced by the combinations of L-OddC and several commonly used chemotherapy drugs in a panel of cultured human cancer cell lines. Growth inhibition resulting from simultaneous exposure to two-drug combinations was determined using the methylene blue staining method. Camptothecin (CPT) and analogs exhibited additives to synergistic interactions with L-OddC by isobologram analysis. These effects were cell type-specific, with the most pronounced synergism being observed in KB oropharyngeal carcinoma and CPT-resistant KB100 cell lines. In KB cells, the total cellular uptake and DNA incorporation of L-OddC were increased by the addition of CPT. One explanation that emerged from enzyme assays of deoxycytidine kinase (dCK) and deoxycytidine monophosphate kinase (dCMPK), key enzymes involved in L-OddC phosphorylation, was that CPT protected against L-OddC-induced reduction in dCK and dCMPK activity. The resulting increase in l-OddC metabolites and incorporation into DNA was associated with enhanced L-OddC cytotoxicity. These findings will be useful in designing future clinical trials of combination chemotherapy with l-OddC and CPT analogs with the potential for a broad use against both hematological and solid tumors.

Airway Sealing Effect and Gastroesophageal Regurgitation of Laryngeal Mask Airway-Proseal in Laparoscopy

Background: The laryngeal mask airway Proseal (LMA-Proseal) is a renewed airway device with a modified cuff to improve the mask seal around the larynx. The classic LMA is known to have an incomplete mask seal during high positive pressure ventilation. Therefore, we examined whether the ventilation via a LMA-Proseal during laparoscopic surgery may be adequate and also observed the complications associated with a LMA-Proseal. Methods: Thirty four patients undergoing elective gynecological laparoscopies were studied. After the insertion of a LMA-Proseal, we measured the peak airway pressure, plateau airway pressure, oropharyngeal leakage pressure, and the leakage around the neck before and after intra-abdominal insufflation. After extubation, we observed the complications, including the gastroesophageal regurgitation using a methylene blue staining method. Results: The peak and plateau airway pressures after pneumoperitoneum increased significantly (P ＜ 0.05), compared with those before insufflation. However, the oropharyngeal leakage pressure was not changed and leakages around the neck were not observed. After extubation, a cuff stained with methylene blue occurred in 10 of 34 patients. Sore throat and dysphagia occurred in 8 and 4 of 34 patients, respectively. Conclusions: A LMA-Proseal during laparoscopic surgery permits adequate airway pressure, but does not protect the gastroesophageal regurgitation.