Blue Dye Method Research Articles

Evaluation of Antitumor Activity of Sivanar Amirtham – A Herbomineral Formulation belonging to Indian Traditional System of Medicine using Dalton's Ascites Lymphoma in Mice

The purpose of the present study is to evaluate the antitumor activity (ATA) of traditional herbal preparation Sivanar Amirtham (SA) on Dalton’s Lymphoma Ascites. Siddha medicine system (SMS) is a traditional system of medicine originated from ancient Tamilakam of South India. Siddha medicine is a traditional healing system from Tamilakam in ancient South India. For our purpose, we have performed acute toxicity (AT) study as per OECD guidelines 423 and ATA by xenograft method. In this study, a single dose of 300, 1000 and 2000 mg/kg of Sivanar Amirtham suspension (SAS) was orally (p.o.) administered in mice and animals were observed for 14 days. For antitumor study (ATS), we have used DAL cells which were intraperitoneally (i.p.) inoculateded into mouse. The ATAs were studied by monitoring the parameters such as cell growth inhibitors, tumor weight measurements, mean survival time of DAL bearing mice as well as changes in depleted haematological and biochemical parameters due to tumorigenesis. The SAS was also evaluated for in vitro cytotoxicity study in different concentration and the viability of cells was determined by exclusion method of trypan blue dye (TBD). The AT study showed no signs of toxicity and no mortality after single administration of SAS. SAS caused significant decrease in packed cell volume (PCV) (value), Tm volume (value) and viable cell count (value), and it prolonged the life span of DAL Tm carrying mice. Haematological and biochemical profiles were reverted to normal levels in SAS treated mice. The results of in vitro cytotoxicity show that SAS showed significant ATA in mice with moderate DAL levels. The IC50 value turned into discovered to be 800 μg/ml from the in vitro cytotoxicity examine. The study strongly suggests that SAS has the potential to be an antitumor medication against DAL cells induced Tm and it can be extrapolated for further cancer (CA) prevention applications.

THE COMPARISON OF RADIOCOLLOID, BLUE DYE AND A COMBINED METHOD FOR SENTINEL LYMPH NODE BIOPSY IN EARLY-STAGE BREAST CANCER

This study aims to compare the performance of radiocolloid, blue dye and a combined method for sentinel lymph node biopsy in early breast carcinoma. One hundred seventy-two clinically node-negative, early-stage breast cancer patients who underwent combined method sentinel lymph node biopsy (SLNB) with 99mTc-nanocolloid and blue dye were recruited. The peri-areolar injection is the method used in both techniques. The hot node contained only radiocolloid, the blue node contained only blue dye, and the hot-blue node contained both tracers. The detection, metastatic, and failure rate of the combined, radiocolloid, and blue dye methods were determined by analysing the number of hot and blue nodes in all patients, all excised nodes, and metastatic nodes. The association between clinicopathological factors and the detection of sentinel lymph nodes (SLN) was analysed. The overall SLN detection rate was 98.2% (169/172) for radiocolloid, 95.9% (165/172) for blue dye, and 99.4% (171/172) for the combined method. The metastatic rate was 19.2% (33/172). Among the 33 metastatic-node cases, the combined, radiocolloid and blue dye method failed to identify 1, 1 and 4 cases, respectively. Therefore, the failure rate of the combined, radiocolloid, and blue dye method was 3.0% (1/33), 3.0% (1/33) and 12.1% (4/33), respectively. No significant association between clinicopathological factors and SLN detection was observed. The combined method of SLNB yielded the highest detection rate when compared to the radiocolloid-alone and blue-dye-alone methods. The radiocolloid method detected more SLN than the blue dye method. Therefore, incorporating the radiocolloid method in the blue dye method will improve the SLN identification rate and reduce the failure rate of metastatic detection.

Application of fluorescence endoscopy with methylene blue dye and indocyanine green dual-tracer method in sentinel lymph node biopsy for women with breast cancer.

Indocyanine green (ICG) allows for the real-time visualization of lymphatic drainage and provides favorable performance for sentinel lymph node (SLN) mapping. However, the limited ability of tissue penetration of the near-infrared fluorescence of ICG may lead to the failure of lymph node detection in the traditional open approach of sentinel lymph node biopsy (SLNB) for breast cancer, especially in overweight or obese patients. To accurately and quickly detect SLNs, we applied fluorescence endoscopy with a dual-tracer method using ICG and methylene blue dye (MBD) in SLNB for breast cancer. We conducted this study to assess the feasibility and application value of this method in minimally invasive surgery. A total of 117 patients who received dual-tracer injection of ICG and MBD prior to endoscopic SLNB from November 2020 to September 2021 were examined in this study. The number of SLNs identified, the SLN identification rate, the time to identify the first SLN, the procedure duration, and the postoperative morbidity were analyzed. Biopsied SLNs could be identified in 116 patients (99.15%) with an average number of 5.12±1.87 per patient. Blue-stained SLNs were found in 99 patients (84.62%) and fluorescent SLNs in 112 patients (95.73%). A total of 34 patients (29.06%) had positive SLNs. In 6 cases (5.13%), the positive SLNs were only stained with ICG fluorescence. In 1 case (0.85%), the positive SLNs were only blue-stained with no fluorescence staining. The mean durations for the identification of the first SLN and endoscopic SLNB were 7.14±6.31 and 37.75±16.94 min, respectively. Upper-limb lymphoedema was observed 5 cases (4.27%) during a median follow-up period of 10 months. The fluorescence endoscopy method assisted by dual tracer facilitates SLN detection with a comparatively short procedure duration and low complication rate. This approach could serve as a new method for SLNB for patients with breast cancer.

Sentinel lypmh node biopsy in early breast cancer: preliminary results of the combined technique of CT lymphography and blue-dye

Aim: The aim of this study was to evaluate the efficacy of CT lymphography in sentinel lymph node biopsy for early stage breast cancer and to investigate its contribution to the conventional blue-dye method.
 Material and Method: A total of 47 patients with early stage breast cancer underwent preoperative CT lymphography for lymph node mapping before sentinel lymph node biopsy with blue-dye method. The lymph nodes identified by CT lymphography and/or blue-dye method were examined for metastatic involvement. The sentinel lymph node detection rates of CT lymphography and blue-dye method were compared using t-tests.
 Results: The sentinel lymph node detection rate with blue-dye method (87.2%) was significantly higher than with CT lymphography (66.0%) (P=0.027). However, the combined method (blue-dye method and/or CT lymphography) increased the detection rate (95.7%) (P=0.267). Benign sentinel lymph nodes were detected more often with CT lymphography (P=0.366), while metastatic sentinel lymph nodes were detected more often with blue-dye method (P=1,000). Upper outer quadrant tumors were detected less successfully with CT lymphography and more successfully with blue-dye method (P=0.220 and P=0.674, respectively). The success rate of CT lymphography in younger patients (less than 50 years old) was higher compared to older patients (P=0.001).
 Conclusion: CT lymphography was found to be insufficient as a standalone method for sentinel lymph node biopsy. However, it could be used as a complementary method to blue-dye method to increase the success of sentinel lymph node detection.

Activation of ERK1/2 by MOS and TPL2 leads to dasatinib resistance in chronic myeloid leukaemia cells.

The development of BCR::ABL1 tyrosine kinase inhibitors (TKIs), such as dasatinib, has dramatically improved survival in cases of chronic myeloid leukaemia (CML). However, the development of resistance to BCR::ABL1 TKIs is a clinical problem. BCR::ABL1 TKI resistance is known to have BCR::ABL1-dependent or BCR::ABL1-independent mechanisms, but the mechanism of BCR::ABL1 independence is not well understood. In the present study, we investigated the mechanism of BCR::ABL1-independent dasatinib resistance. The expression and activation level of genes or proteins were evaluated using array CGH, real time PCR, or western blot analysis. Gene expression was modulated using siRNA-mediated knockdown. Cell survival was assessed by using trypan blue dye method. We found that dasatinib-resistant K562/DR and KU812/DR cells did not harbour a BCR::ABL1 mutation but had elevated expression and/or activation of MOS, TPL2 and ERK1/2. In addition, MOS siRNA, TPL2 siRNA and trametinib resensitized dasatinib-resistant cells to dasatinib. Moreover, expression levels of MOS in dasatinib non-responder patients with CML were higher than those in dasatinib responders, and the expression of TPL2 tended to increase in dasatinib non-responder patients compared with that in responder patients. Our results indicate that activation of ERK1/2 by elevated MOS and TPL2 expression is involved in dasatinib resistance, and inhibition of these proteins overcomes dasatinib resistance. Therefore, MOS, TPL2 and ERK1/2 inhibitors may be therapeutically useful for treating BCR::ABL1-independent dasatinib-resistant CML.

Neuroprotective effects of chloroquine on neurological scores, blood-brain barrier permeability, and brain edema after traumatic brain injury in male rats

Background: Traumatic brain injury (TBI) is one of leading causes of death among young people worldwide. Chloroquine, an antimalarial drug, has been shown to easily cross the blood-brain barrier (BBB) and inhibit autophagy in a variety of disorders, including Alzheimer disease and brain ischemia. We investigated the effects of chloroquine on neuronal protection after induction of brain trauma in male rats.Methods: A total of 120 male Wistar rats were treated with chloroquine at doses of 1.5, 3, and 6 mg/kg intraperitoneally after induction of diffuse TBIs. The veterinary coma scale was used to assess short-term neurological deficits. BBB disruption was evaluated using the Evans Blue dye method 6-hour post-injury. Vestibulomotor function was evaluated using the beam walk and beam balance methods. Histopathological changes in the brain tissue in different groups were evaluated using light microscopy and hematoxylin-eosin staining. Brain water and cerebrospinal fluid (CSF) contents of matrix metalloproteinase 9 (MMP-9) were assessed using the wet/dry method and enzyme-linked immunosorbent assay, respectively.Results: The results showed that injecting chloroquine (3 and 6 mg/kg) 30 minutes after TBI significantly reduced brain edema and BBB disruption, and recovered neurological deficits post-TBI (P<0.01). Furthermore, CSF MMP-9 was significantly reduced after administration of 1.5 mg/kg chloroquine (P<0.01).Conclusion: Chloroquine has neuroprotective effects in the brain, and thus, has the potential to mitigate the effects of brain trauma. It is possible that the anti-inflammatory and neurogenic effects of chloroquine are due to a decrease in MMP secretion in the CSF.

Sentinel Node Identification with Hybrid Tracer-guided and Conventional Dynamic Sentinel Node Biopsy in Penile Cancer: A Prospective Study in 130 Patients from the Two National Referral Centres in Sweden

BackgroundStudies suggest that a hybrid indocyanine green (ICG)-99mTc-nanocolloid tracer improves sentinel node (SN) identification compared to conventional dynamic sentinel node biopsy (DSNB). ObjectiveTo investigate hybrid tracer–guided SN identification in a multicentre setting and determine false-negative (FN) and complication rates. Design, setting, and participantsA total of 130 patients with penile cancer scheduled for DSNB were prospectively included between February 2016 and December 2017 at two national Swedish referral centres. ICG-99mTc-nanocolloid hybrid tracer was used in the standard DSNB protocol. InterventionSNs were identified intraoperatively using radioguidance, fluorescence imaging, and blue dye. Outcome measurements and statistical analysisThe number of SNs identified by each tracer and the rates of complications and nodal recurrence during median follow-up of 34 mo were recorded. Differences in proportions between groups were compared using χ2 and McNemar’s tests. Results and limitationsOverall, 453 SNs were identified preoperatively via single-photon emission computed tomography/computed tomography. Among the 425 SNs excised, radioguidance, fluorescence, and blue dye identified 414 (97%), 363 (85%), and 349 (82%), respectively. Fluorescence imaging helped to detect six SNs that were negative using the other tracers, two of which were from the same patient and contained metastases. Histopathological examination detected 33 metastatic SNs in 20/130 patients (15%). The FN rate was 12% per groin (95% confidence interval 8–16%). ConclusionsIdentification of SNs in patients with penile cancer relies mainly on radioguidance, while fluorescence (ICG) and blue dye methods for optical SN identification are comparable. However, the value of fluorescence imaging should be further evaluated in studies with long-term follow-up. Patient summaryIn this study, we investigated addition of a dye called indocyanine green (ICG) for assessment of lymph nodes in patients with cancer of the penis. ICG did not improve the rate of detection of nodes most likely to harbour cancer because of their location in the drainage pathway for lymphatic fluid, but did help in identifying additional metastases.

TiO2 ceramic membrane decorated with Fe3O4–Ag composite nanoparticles for produced water treatment

This study focused on the surface modification of commercial TiO2 membranes with Fe3O4 decorated silver (Ag) nanoparticles (Fe3O4–Ag) via chemical attachment. Firstly, the Ag concentration on Fe3O4 was optimized, and different composites were prepared and characterized. Secondly, the optimal composite was used to prepare novel TiO2/Fe3O4–Ag ceramic membranes via surface coating through tetraethyl orthosilicate (TEOS) crosslinking. The membranes were characterized using SEM, EDX, FTIR, XRD, and contact angle. Biofouling resistance of the membranes was investigated using the Coomassie Blue dye method. The coated membranes were tested for water flux, chemical oxygen demand (COD) rejection, and biofouling resistance. Results showed that all coated membranes exhibited higher water flux. For example, the membrane with a 1.25 wt% Fe3O4–Ag coating showed the highest filtration flux of 1445 L/m2h (LMH) compared to the pristine membrane (379 LMH) without compromising the COD rejection. The resistance of the membrane to biofouling increased with the increase of Fe3O4–Ag nanoparticle concentration. The obtained results demonstrate the great potential of TiO2/Fe3O4–Ag ceramic membranes for the treatment of produced water.

A 2A ADENOSINE RECEPTORS REGULATE MULTIPLE ORGAN FAILURE AFTER HEMORRHAGIC SHOCK IN MICE.

Trauma hemorrhagic shock (T/HS) is a clinical condition that causes multiple organ failure that needs rapid intervention. Restricted oxygen at the cellular level causes inflammation and subsequent cell death. Adenosine triphosphate is the universal intracellular energy currency and an important extracellular inflammatory signaling molecule. Adenosine, an endogenous nucleotide formed as a result of the breakdown of adenosine triphosphate, is also released during T/HS. Adenosine binds to four G protein-coupled receptors (A 1R , A 2a , A 2b , A 3R ) called adenosine receptors or P1 receptors. In the present study, we evaluated the effect of activation, inactivation, and genetic absence of A2aR (A2aR -/- mice) on T/HS-induced multiple organ failure. Wild-type mice were pretreated (30 min before shock induction) with an agonist or antagonist and then subjected to T/HS by withdrawing arterial blood and maintaining the blood pressure between 28 and 32 mm Hg. A2aR -/- mice were subjected to T/HS in the absence of pharmacologic treatment. Neutrophil sequestration was assessed by detecting myeloperoxidase, and Evans blue dye (EBD) method was used to analyze lung permeability. Blood and lung inflammatory cytokine levels were determined by sandwich enzyme-linked immunosorbent assay. The liver enzymes aspartate aminotransferase and alanine aminotransferase were determined spectrophotometrically from plasma. Activation of the apoptotic cascade was evaluated using a mouse apoptosis array. Our results demonstrate that the selective A2aR agonist CGS21680 decreases lung neutrophil sequestration, lung proinflammatory cytokines IL-6 and TNF-α, and bronchoalveolar lavage EBD. Pretreatment with the selective antagonist ZM241385 and genetic blockade in A2aR -/- mice increased neutrophil sequestration, proinflammatory cytokine levels, and bronchoalveolar lavage fluid EBD. The myeloperoxidase level in the lung was also increased in A2aR -/- mice. We observed that antiapoptotic markers decreased significantly with the absence of A2aR in the lung and spleen after T/HS. In conclusion, our data demonstrate that activation of A2aR regulates organ injury and apoptosis in the setting of T/HS.

Robotic device for transcranial focussed ultrasound applications in small animal models.

Focussed Ultrasound (FUS) combined with microbubbles (MBs) was proven a promising modality for non-invasive blood brain barrier disruption (BBBD). Herein, two devices for FUS-mediated BBBD in rodents are presented. A two-axes robotic device was manufactured for navigating a single element FUS transducer of 1MHz relative to the brain of rodents. A second more compact device featuring a single motorized vertical axis was also developed. Their performance was assessed in terms of motion accuracy, MRI compatibility and trans-skull BBBD in wild type mice using MBs in synergy with pulsed FUS. Successful BBBD was evidenced by the Evans Blue dye method, as well as by Fibronectin and Fibrinogen immunostaining. BBB permeability was enhanced when the applied acoustic intensity was increased. The proposed devices constitute a cost-effective and ergonomic solution for FUS-mediated BBBD in small animal models. Further experimentation is needed to examine the repeatability of results and optimise the therapeutic protocol.

Detailed analysis of structural, optical and photo catalytic properties of spinel nickel doped magnesium zinc ferrites at different substitutions

Spinel nickel doped magnesium zinc ferrite nanoparticles under different substitutions have been prepared using co-precipitation technique. The characterization studies which include crystallite size, crystalline solid structure and optical properties of the prepared nanoparticles were examined using x-ray diffraction (XRD), fourier transmission infrared spectrum (FTIR), scanning electron microscope (SEM) and absorbance that was measured by ultraviolet visible (UV) spectroscopy. As increasing the Ni concentration, the optical band gap increases from 3.32 eV to 4.57 eV. In this research work, the photocatalytic assessment of Ni-added-Mg-Zn nano-ferrites has been investigated by using bulb light degradation (methylene blue dye (60 W) method. Mg0.5Zn0.5-xNixFe2O4 spinel nano-ferrites (x = 0.125, 0.250, 0.375) were produced by co-precipitation method. The photocatalytic study has been reported by analyzing the degradation of methylene blue dye under sunlight irradiation. The better-quality photocatalytic activity with increased Ni content was investigated for the samples under study for a 40-minute exposure of × = 0.125, a maximum degradation efficiency of 61 precent was acquired. The phase-change was appeared due to shifting of peaks to the higher angle in Mg-Zn ferrite samples as a result of increments in Ni contents. According to vegard's law, the lattice parameter (a) of the crystal increases with increasing the concentration of nickel ion. Phase identification of magnesium-zinc ferrite nanoparticle was done by using X-ray diffraction (XRD).

Sentinel lymph node biopsy using methylene blue dye and intraoperative palpation method in node-negative early breast cancer

Background: Early breast cancer with clinical or radiological node-negative axilla is treated with breast conservation surgery where lumpectomy with axillary dissection is done. Sentinel lymph node biopsy (SLNB) is an acceptable alternative to axillary clearance and has relatively lesser morbidity. SLNB methods include radioisotope (RI)-guided, blue dye-guided, or a combination of both. However, access to RI can be limited in certain geographic locations. Objective: This study aimed to determine the effectivity of methylene blue (MB) dye-guided SLNB as an alternative to RI. Methods: In this investigation, 43 clinically nodenegative early breast cancer patients were prospectively enrolled. SLNB was performed using MB dye (1%) administered to the peritumoral or periareolar region. The histopathology reports of the harvested nodes were studied, and the results were computed using SPSS and 2×2 contingency table. Results: The sensitivity, specificity, false-negative rate (FNR), and accuracy of MB-guided SLNB in our study were 92.8%, 100%, 7.14%, and 97.7%, respectively. Conclusion: The use of MB dye along with intraoperative palpation after meticulous lymph node dissection in each level is more effective and has lower FNR than RI.

STUDY THE SPECIFICATIONS OF ACTIVATED CARBON PREPARED FROM AVOCADO SEEDS USING CARBONATION AND CHEMICAL ACTIVATION

Activated carbon was prepared from Avocado Seeds using different percentages of sodium hydroxide and under the same conditions. Through the study, it was found that increasing the base leads to the production of new good varieties of a neutral nature. The study proved through physical and chemical examinations of the prepared samples that gave the best results when compared with other samples for activated carbon produced by well-known companies, the most important tests conducted for the prepared samples are the determination of the density, the iodine number (IN), the low ash content, the measurement of the external surface area by the adsorption method of the methylene blue dye from its aqueous solution, the measurement of moisture and ash content.

Antitubercular study on stem bark of Albizia procera (ROXB.) BENTH using Microplate Alamar Blue assay (MABA)

Albizia procera commonly known as white siris is a large deciduous tree of the family Mimosideae, widely distributed throughout India. Traditionally the plant is used in convulsions, pain, delirium, cancer and septicemia. The decoction of bark is given for rheumatism, hemorrhage and is considered useful in treating pregnancy problems, for stomach ache and sinus.The preliminary phytochemical screening of ethanol extract of Albizia procera shows the presence of active constituents like flavonoids, phenolic compounds and tannins. Earlier studies show the flavonoids and phenolic compounds play major role in antitubercular activity. Based on this, we made an attempt to evaluate the antitubercular potential of this plant. The ethanolic extract and its fractions (Hexane and Ethyl acetate) were investigated for antitubercular activity using Alamar Blue dye method (MABA), in which Mycobacterium tuberculosis was sensitive upto 3.12μg/ml.The extract and fractions show better anti tubercular activity than streptomycin and potentially equal to pyrazinamide and ciprofloxacin. The observed activity may be due to the presence of flavonoids and phenolic compound which is confirmed by HPTLC. This investigation on stem bark of Albizia procera has the potential to be developed further into a natural Anti-TB drug.

Acute effects of estradiol-17β on plasma volume and uterine cell proliferation in sheep

Ovarian steroids play an important role in increasing plasma volume in pregnant females and preparing the uterus for implantation. We hypothesized that a short duration of increased estradiol-17β (E2) would increase plasma volume and uterine cell proliferation in ovariectomized ewes. Adult non-pregnant Romanov ewes (n = 15) were ovariectomized. After ovariectomy, ewes were individually housed and were offered water at ad libitum intake and were fed a pelleted diet at maintenance once daily according to body weight. After at least 30 days post-ovariectomy ewes were fasted and received an implant placed in the axillary region that contained 100 mg of E2 (E2; n = 8) or a sham implant with no E2 (CON, n = 7). After 24 h, ewes were weighed prior to plasma volume measurement procedures. Plasma volume was determined using the Evans blue dye method. Blood samples were taken at 0 (pre dye injection), 5, 10, 15, 20, 25, 30, 40, 50 and 60 min after dye injection. After the final blood collection, ewes were euthanized with an overdose of sodium pentabarbital and uterine weights were recorded. Uterine cross-sections were fixed in formalin for immunohistochemical localization of Ki67 (a marker of proliferating cells) followed by image generation of luminal epithelium and endometrial stroma (5 areas each/tissue section) and analysis to determine the proportion of proliferating cells. Plasma volume tended to be greater in E2 vs CON (2.75 ± 0.11 vs. 2.54 ± 0.12 L, P = 0.07) and uterine weights were greater in E2 vs CON (27.25 ± 2.35 vs. 17.35 ± 2.51 g, P < 0.01). Water intake after implant placement was similar in E2 and CON (3.85 vs. 4.87 ± 0.67 L; P = 0.28). Cell proliferation in the luminal epithelium was greater in E2 vs CON (6.55 vs. 1.2 ± 1.75%, P = 0.02) and stromal cells tended to be greater in E2 vs CON (0.59 vs 0.37 ± 0.06%, P = 0.07). Our results demonstrate that E2-treatment tends to increase plasma volume acutely and increases uterine cell proliferation in ewes.

The combination of contrast-enhanced ultrasonography with blue dye for sentinel lymph node detection inclinically negative node breast cancer.

The aim of this prospective study was to evaluate the value of the combination of contrast-enhanced ultrasonography (CEUS) and blue dye (BD) for SLN detection in patients with clinically negative node breast cancer. Patients with clinically negative node breast cancer were randomized into two cohorts for SLN biopsy (SLNB): the combination method cohort using CEUS and BD together, and the single BD method cohort. Standard axillary lymph node dissection was performed if any of the SLNs confirmed positive by pathology. The identification rate, the number of SLNs removed and recurrence-free survival (RFS) rates were evaluated between two cohorts. In addition, we assessed the sensitivity, specificity, accuracy, false-negative rate of CEUS for diagnosis of SLNs based on patterns of CEUS enhancement. 144 consecutive patients with clinically negative node breast cancer were randomized into two cohorts. Each cohort consisted of 72 cases. In the combination method cohort, contrast-enhanced lymphatic vessels were clearly visualized and SLNs were accurately localized in 72 cases. The identification rate and the mean number of SLNs detected by the combination method were 100% (72/72) and 3.26 (1-9), respectively. In contrast, in the single BD method cohort, SLNs in 69 cases were successfully identified. The identification rate and the mean number of SLNs using BD alone were 95.8% (69/72) and 2.21 (1-4), respectively. According to patterns of CEUS enhancement, the sensitivity, specificity, accuracy, and the FNR of CEUS for SLN diagnosis were 69.2%, 96.6%, 91.7%, and 30.8%, respectively. After a median follow-up of 50months for the combination method cohort and 51months for the blue dye alone cohort, five patients in the combination method cohort and nine in the blue dye alone cohort had recurrence. RFS rates showed no significant difference (P = 0.26) between two cohorts. The combination of CEUS and BD is more effective than BD alone for SLNB in clinically negative node patients with an identification rate as high as 100%. Use of BD and CEUS in combination may provide the possibility of a non-radioactive alternative method for SLNB in centers without access to radioisotope.

Sentinel Lymph Node Biopsy in Early Breast Cancer Using the Indocyanine Green Fluorescence Technique: A Validation Study

INTRODUCTION: Introduction: Sentinel lymph node biopsy (SLNB) is now recognized as the standard of care for early breast cancer patients with negative axillary lymph nodes. Various approaches for Sentinel Lymph Node (SLN) identification using either the blue dye method or scintigraphy alone or their combination have been proposed. However, this method is costly and may not be applicable in certain developing countries. SLNB involving the use of indocyanine green (ICG) offers several advantages, and it is valid and safe when in direct comparison with the blue dye method and scintigraphy. Hence, we performed SLNB using this method in early breast cancer as the first center that involves the use of ICG in Malaysia. We performed validation study on this method with the aims to determine its sensitivity and safety profile. MATERIALS AND METHODS: This is a validation and non-randomised prospective observational study involving 20 patients underwent SLNB wherein ICG is used for localisation. The patients were recruited according to the recommendations stipulated in the Malaysia Clinical Practice Guideline. RESULT: The average number of SLNs removed per patient was 4.0 (range, 3–6) with sentinel lymph nodes detection rate at 98.75% (79/80). The false negative rate is at 5%. No adverse events were observed in all cases. CONCLUSION: The ICG fluorescence method is simple, reliable and safe. Moreover, it demonstrates a high SLN detection rate with a low false-negative rate, and it does not require a special instrument for radioisotope use.

Development and Validation of Direct Spectrophotometric Method for the Estimation of Glimepiride

Aims: To develop simple, sensitive and direct spectrophotometric methods for the estimation of widely prescribed antidiabetic hypoglycemic agent Glimepiride in pure and pharmaceutical dosage form.&#x0D; Place and duration of study: Research Centre of Taywade Group of Institution Koradi, Nagpur affiliated to Rashtrasant Tukadoji Maharaj Nagpur University Nagpur between June 2020 and March 2021.&#x0D; Methodology: Two spectrophotometric methods were developed based on ion-pair formation of drug with Cresol Red dye (Method A) and Bromophenol Blue dye (Method B) in methanol and chloroform. The possible reaction mechanism was proposed with evaluation of statistical parameters. The methods were validated for its application to determine Glimepiride in bulk as well as in pharmaceutical formulations&#x0D; Results: The Beer's law was found linear in the range 10 - 60 µgml-1 at 450 nm for method A and 2 - 20 µgml-1 at 578 nm for method B, respectively. The linear regression equation obtained by applying least square regression analysis for Glimepiride were found to be Absorbance = 0.0136 x Concentration in µgml-1 + 0.028; R2 = 0.9965 for method A and Absorbance = 0.0428x Concentration in µgml-1 +0.0722; R2 = 0.9949 for method B. The Sandell's sensitivity was found to be 0.0696 and 0.0177 for method A and B respectively. The apparent molar absorptivity calculated to be for both methods were 6.6724 x103 and 2.0999 x104 l mol-1 cm-1.&#x0D; Conclusion: Two direct spectrophotometric methods for determination of Glimepiride have been developed successfully and validated for sensitivity, accuracy, precision, repeatability and robustness as per ICH guidelines. The developed methods are suitable for the routine estimation of Glimepiride in pure and dosage form.

Protection Against Blood-Brain Barrier Permeability as a Possible Mechanism for Protective Effects of Thymoquinone Against Sickness Behaviors Induced by Lipopolysaccharide in Rats

Background: Blood-brain barrier (BBB), as well-known protection for the brain, plays an active role in normal homeostasis. It might be changed by a range of inflammatory mediators to have a role in sickness behaviors. Objectives: Regarding the anti-inflammatory effects of thymoquinone (TQ), its protection against BBB permeability, as a possible mechanism for protective effects against sickness behaviors elicited by lipopolysaccharide (LPS), was evaluated in rats. Methods: The animals were grouped as follows and treated (n = 10 in each): (1) control (saline); (2) LPS 1 mg/kg, was injected two hours before behavioral tests for two weeks; (3-5) 2, 5, and 10 mg/kg TQ, respectively was injected 30 min before LPS injection. Open-field (OF), elevated plus-maze (EPM) and Forced Swimming test (FST) were done. Finally, the animals were anesthetized to evaluate for BBB permeability using Evans blue (EB) dye method. Results: Compared with control, LPS decreased the peripheral distance and crossing and also total crossing and distance in OF, (P &lt; 0.01 - P &lt; 0.001). The central crossing and distance and central time in all three treatment groups were more than LPS (P &lt; 0.05 - P &lt; 0.001). LPS also reduced the entries and the time spent in the open arm while increased the time spent in the closed arm in EPM (P &lt; 0.05 - P &lt; 0.001). The effects of LPS were reversed by TQ (P &lt; 0.05 - P &lt; 0.001). In FST, the immobility time and active time were increased and decreased by LPS compared with control (P &lt; 0.001), respectively. In all three TQ-treated groups, the active and climbing times were more while the immobility time was fewer than the LPS (P &lt; 0.05 - P &lt; 0.001). The animals of the LPS group showed more EB dye content in their brain tissue than the control group (P &lt; 0.05 - P &lt; 0.001). TQ significantly reduced EB dye content of the brain tissues (P &lt; 0.05 - P &lt; 0.001). Conclusions: According to this study, protection against BBB permeability as a possible mechanism for the protective effects of TQ against sickness behaviors induced by LPS might be suggested.

Blue poo: impact of gut transit time on the gut microbiome using a novel marker

Background and aimsGut transit time is a key modulator of host–microbiome interactions, yet this is often overlooked, partly because reliable methods are typically expensive or burdensome. The aim of this...