Intravesical combination therapies for non-muscle invasive bladder cancer: Recent advances and future directions.

Bladder cancer is the 10th most frequently diagnosed cancer worldwide with 5-year survival rate around 70%. The current first-line treatment for non-muscle invasive bladder cancer is transurethral resection of bladder tumours followed by intravesical Mycobacterium Bovis Bacillus Calmette-Guérin (BCG) immunotherapy. However, tumor recurrence rate is still high ranging from 31% to 78% within five years. To avoid radical cystectomy, intravesical combination therapies have been developed as salvage treatments to overcome BCG failure. Recent advances in diagnostics thanks to tumor molecular profiling and in treatment such as development of immunotherapies provides more treatment options beyond BCG treatment. This also goes hand-in hand with formulation advances to deliver these new therapies where traditional drug delivery systems might not be suitable, which in turn is completed by challenges to deliver drugs via the intravesical route. In this article the aim was to provide an in-depth analysis of the current developments of intravesical combination therapies, ranging from relatively simple combinations of mixing existed intravesical therapeutic agents (immunotherapies and chemotherapies) to the combined formulations containing advanced gene therapies and targeted therapies, with special focus on therapies that have made it to the clinical trial stage. In addition, recent attempts to utilize device-assisted treatments and novel drug delivery platforms are included. This review also highlights the limitations that still need to be overcome such as the inadequate studies on newly explored drug carriers and proposes potential directions for future work to overcome BCG-failure.

The first evaluation of the effectiveness of canine vaccination schedule by two commercial vaccines in Iran.

BACKGROUND: Canine Parvovirus type 2 (CPV-2) is a member of the Parvoviridae family with a global distribution and causes pathogenicity in puppies aged from 6 weeks to 6 months. It should be noted that Maternally Derived Antibodies (MDA) have protection against CPV-2 in the first weeks of puppies' life. However, MDA declines with age. The most important influential factor is timely vaccination against CPV-2. METHODS: In this study, 24 healthy 8-week-old terrier puppies were selected and divided into three identical groups based on a randomized, double-blind comparative trial. One of which was called the control group that was injected with the physiological serum. The second group was the group A that was vaccinated by the vaccine provided by Biocan DHPPi+L (Bioveta, Czech). The third group was group B that was vaccinated by the vaccine of Duramune Max 5 + LCI / GP (Fort Dodge Animal Health, USA) from 8 to 16 weeks of their life at every 4 weeks. Then serum samples were analyzed with HI and ELISA tests. RESULTS: The MDA titer was protective in some puppies until 18 weeks of age. Also, after the first vaccination, all puppies had a protective titer against CPV-2, and Duramune vaccine had seroconverted after the first injection and Biocan had seroconverted after the second injection. CONCLUSIONS: It is recommended that to reduce the risk of vaccine failure: such as the MDA titer should be measured in puppies before designing a vaccination schedule.

Current Topical Strategies for Skin-Aging and Inflammaging Treatment: Science versus Fiction.

Aging is a natural phenomenon that affects the whole body, including the skin. As we age, endogenous and exogenous factors cause our skin to become thinner, paler, and wrinkled. Although the underlying mechanisms of the pathogenesis of skin aging are not entirely known, multiple pathways have been proposed. Inflammaging has recently emerged as a pathway that correlates aging and age-related diseases with inflammation. This review discusses the role and pathways of inflammaging that lead to skin aging. Moreover, strategies and current topical approaches for skin-aging treatment are discussed. Studies over the past 10 years suggested that DNA damage and oxidative stress are the most critical mechanisms in skin aging, and both are interlinked with inflammaging. Several treatments for skin aging have been considered such as antioxidants, hormone replacement therapy, and vitamins. To deliver anti-aging agents topically, researchers adopted numerous approaches to enhance skin penetration including physical, chemical, or biomaterial enhancers and carrier-based formulations. In recent years, consumers' demands for anti-aging products have considerably risen, leading to robust growth in the anti-aging market. Therefore, further in-depth studies are necessary to understand skin-aging mechanisms and evaluate the efficacy of anti-aging products to protect consumers worldwide by providing them safe and effective over-the-counter skin-aging formulations.

Immunohistochemical detection of V600E BRAF mutation is a useful primary screening tool for malignant melanoma.

BACKGROUND: We compared the use of an immunohistochemical (IHC) method using a monoclonal antibody to BRAF V600E (which detects the main BRAF mutation) with existing DNA probe screening in tissue samples from 71 patients with malignant melanoma. MATERIALS AND METHODS: Paraffin blocks were cut to provide consecutive slides for haematoxylin and eosin staining, and for known positive micro-array DNA control material. IHC was performed by the Optiview detection system. All slides were scored independently by the clinical lead and the laboratory lead using a positive/negative system. RESULTS: The DNA method found 26 samples to be positive, the IHC found 21 to be positive, giving a sensitivity value for IHC of 80.8%. However, all of the 45 samples found to be negative by DNA were also negative by IHC, giving a specificity of 100%. There were 66 instances of full agreement, giving a concordance of 93%. Together, these data give a kappa statistic of 0.843, indicating very good agreement. CONCLUSION: The data reveal a very close link between the two methods, supporting the use of the V600E as a primary screen for BRAF mutations in malignant melanoma. Samples found to be negative by this method may be retested by the DNA probe method. IHC detection conserves patient DNA from tumour blocks as only one section is required to perform the assay. The V600E antibody method is considerably cheaper and faster than the DNA probe assay, with a turn-around time of 24-48 hours, enabling more rapid clinical management.

Identifying avoidable switchboard delays in England's NHS hospitals: phase one of the national SWITCH project.

BACKGROUND: Inter-hospital communication frequently requires mediation via a switchboard. Identifying and eliminating switchboard inefficiencies may improve patient care. METHODS: All 175 acute hospital switchboards in England were contacted six times. Call contents and duration were recorded. No clinician calls or bleeps were connected. RESULTS: The mean delay before contacting a switchboard operative was 55±46 seconds. 115 hospitals (66%) used automated switchboards; 34 of these (30%) had infection control messages. Robot operators introduced an additional 40 second delay versus humans (mean 70.3±28 versus 29.8±23 seconds, p<0.0001). Multivariate analysis identified robot operators (HR 5.1, p<0.0001) and infection control messages (HR 2.9, p=0.003) as predictors of delays over 60 seconds. CONCLUSIONS: There are significant avoidable delays in contacting switchboard operatives across England. Quality improvement is underway.

Elaboration of polymersomes versus conventional liposomes for improving oral bioavailability of the anticancer flutamide.

AIM: Flutamide is an outstanding anticancer drug with poor oral bioavailability. This is the first work to investigate the potential of polymersomes versus conventional liposomes to improve flutamide bioavailability. MATERIALS & METHODS: Polymersomes were prepared by solvent-switching technique and successfully optimized with excellent nanometric size (143 nm) and ζ-potential (-33.4 mV). Physicochemical characterization, stability in gastrointestinal tract and in vivo oral pharmacokinetics in male Sprague-Dawely rats were performed. RESULTS: A significantly higher stability in simulated intestinal fluid was demonstrated by polymersomes compared with liposomes. Great improvement in flutamide oral bioavailability in polymersomes compared with both liposomes and drug suspension was obtained. CONCLUSION: Polymersomes are promising nanoplatforms to overcome stability problems of liposomes and to improve flutamide oral bioavailability.

New embedding and staining systems PrestoCHILL and Presto stainer for application in the advancement of Mohs micrographic surgery.

BACKGROUND: Mohs micrographic surgery (MMS) involves evaluation of frozen tissue sections to determine complete circumferential and deep tissue margin clearance of skin tumours. PrestoCHILL and Presto stainer devices are two new innovative tools which bring benefits of automation, speed and efficiency to the preparation of frozen section analysis in MMS. The devices were assessed at Viapath's Tissue Science Mohs laboratory at Guy's Cancer Centre. MATERIAL AND METHODS: A total of 279 samples from 10 anatomically different facial sites. These included nose (95), lip (24), forehead (47), cheek (25), eyelids (34), temple (9), chin (15), ear (17), scalp (6) and neck (7). These were analysed using both devices simultaneously. RESULTS: The PrestoCHILL device was measured for accuracy of tissue orientation by determining how many of the cases examined microscopically had complete margin and full epidermis preservation. The precision and reproducibility of the Presto stainer was evaluated by the consistency of achieving ideal standards of staining quality as defined by the department's internal quality control check, on stained sections examined and evaluated microscopically. The mean (standard deviation) score for accuracy for the PrestoCHILL across all tissue facial sites was 93.5 (11)%; the mean (standard deviation) score for precision/reproducibility of the Presto stainer was 96.5 (11)% (both p < 0.05). CONCLUSION: The devices combined offer an assured accuracy and precision performance, which is reproducible across all facial tissue types examined. The devices represent a key step forward in the introduction of improved automated embedding and staining procedures within MMS.

A multicentre study of the precision and accuracy of the TruSlice and TruSlice Digital histological dissection devices.

BACKGROUND: Five key factors enabling a good surgical grossing technique include a flat uniformly perpendicular specimen cutting face, appropriate immobilisation of the tissue specimen during grossing, good visualisation of the cutting tissue face, sharp cutting knives and the grossing knife action. TruSlice and TruSlice Digital are new innovative tools based on a guillotine configuration. The TruSlice has plastic inserts whilst the TruSlice Digital has an electronic micrometre attached: both features enable these dissection factors to be controlled. The devices were assessed in five hospitals in the UK. MATERIAL AND METHODS: A total of 267 fixed tissue samples from 23 tissue types were analysed, principally the breast (n = 32) skin (30), rectum (28), colon (27) and cervix (17). Precision and accuracy were evaluated by measuring the defined thickness, and the consistency of achieving the defined thickness of tissue samples taken respectively. Both parameters were expressed as a total percentage of compliance for the cohort of samples accessed. RESULTS: Overall, the mean (standard deviation) score for precision was 81 (11) % whilst the accuracy score was 82 (11) % (both p < 0.05, chi-squared test), although this varied with type of tissue. Accuracy and precision were strongly correlated (rp = 0.83, p < 0.001). CONCLUSION: The TruSlice Digital devices offer an assured precision and accuracy performance which is reproducible across an assortment of tissue types. The use of a micrometre to set tissue slice thickness is innovative and should comply with laboratory accreditation requirements, alleviating concerns of how to tackle issues such as the 'measurement of uncertainty' at the grossing bench.

Development of new and accurate measurement devices (TruSlice and TruSlice Digital) for use in histological dissection: an attempt to improve specimen dissection precision.

Histological dissection of human tissue has relied on conventional procedures, which have largely remained unchanged for decades. Practices to determine measurement parameters employed in these procedures have largely relied on the use of rulers and weighing scales. It is well documented in the scientific literature that both fixation and processing of tissue can significantly affect the viability of the of tissue sections both for tinctorial and immunocytochemical investigations. Both of these factors can be compounded in their negative effects by inappropriate sampling of tissue at histological cut up. There are five key factors to ensure good surgical grossing technique, flat uniformly perpendicular specimen cutting face, appropriate immobilisation of the tissue specimen during grossing, good visualisation of the cutting tissue face, sharp cutting knives and the grossing knife action. Meeting these factors implies the devices are fit for purpose. Here we describe an innovative approach to designing cut up devices to improve accuracy and precision, which take these five key requirements into consideration. The devices showed accuracy and precision, enabling tissue slices to be produced in a uniformly perpendicular fashion to within 2 mm in thickness and to enable consistency and reproducibility of performance across a series of tissue types. The application of a digital rule on one of these devices ensures accuracy and also enables quality control issues to be clearly assessed. As cellular pathology laboratories conform to ever increasing standards of compliance and performance in practice, the advent of assured precision and accuracy at cut up is awaited. Recommendations from accreditation bodies such as the United Kingdom Accreditation Service (UKAS) continue to push for improvements in this area of histological investigation. These newly designed devices may give the answers to these requirements and provide the impetus for a new generation of innovative equipment for histological dissection.

Pan-cytokeratin markers for rapid frozen section immunocytochemistry from head and facial Mohs cases of basal cell carcinoma: a comparison and evaluation to determine the marker of choice.

The application of immunocytochemistry in the field of Mohs micrographic surgery (MMS) is well established. This study evaluates the use of pan-cytokeratins (AE1/AE3, MNF116 and AE1/AE3+PCK26) in the assessment of basal cell carcinoma (BCC) on frozen tissue debulk specimens. Fifty-five cases of BCC, all from head and facial sites, were assessed in the study. In addition to staining all cases for the three cytokeratin antibodies under investigation, sections were also stained with haematoxylin and eosin (H&E) to demonstrate tumour architecture and morphology. All sections for immunocytochemistry were stained on a Roche Ventana BenchMark Ultra automated platform employing a rapid frozen section protocol. Results were assessed based on the intensity of staining of keratinocytes (scale: 0-100%), as well as sensitivity of staining determined by the total percentage of keratinocytes stained within the tissue section. AE1/AE3 demonstrated the most consistent staining both in terms of intensity of staining and sensitivity, with a mean of 99.1% and 99.9%, respectively. AE1/AE3+PCK26 average results indicated scores of 70.6% for intensity and 87.2% for sensitivity, with MNF116 scoring 92.9% for intensity but only 57.3% for sensitivity. The data indicate that AE1/AE3 is the best pan-cytokeratin antibody to use in the assessment of BCC in MMS. The use of cytokeratin immunocytochemistry is justified in morphologically complex cases of BCC, or in cases where dense inflammatory infiltrate surrounding any suspicious cells make identification of small numbers of tumour cells difficult to determine with just an H&E stain. The significant rationale is that cytokeratin staining is a valuable adjunct in the study of tumour cell assessment in cases of MMS for BCC. In addition, the use of anti-AE1/AE3 cytokeratin antibodies provides the most consistent staining results for such cases.

Oral tremor induced by galantamine in rats: a model of the parkinsonian side effects of cholinomimetics used to treat Alzheimer's disease.

Anticholinesterases are the most common treatment for Alzheimer's disease, and, in recent years, a new group of cholinesterase inhibitors (i.e. rivastigmine, galantamine, and donepezil) has become available. Although these drugs improve cognitive symptoms, they also can induce or exacerbate parkinsonian symptoms, including tremor. The present studies were conducted to determine if galantamine induces tremulous jaw movements, a rodent model of parkinsonian tremor, and to investigate whether these oral motor impairments can be reversed by co-administration of adenosine A(2A) antagonists. The first experiment demonstrated that systemic injections of galantamine (0.75-6.0 mg/kg I.P.) induced a dose-related increase in tremulous jaw movements in rats. In a second study, co-administration of the muscarinic antagonist scopolamine (0.0156-0.25 mg/kg I.P.) produced a dose dependent suppression of tremulous jaw movements induced by a 3.0 mg/kg dose of galantamine, indicating that galantamine induces these tremulous oral movements through actions on muscarinic acetylcholine receptors. In two additional studies, analyses of freeze-frame video and electromyographic activity recorded from the lateral temporalis muscle indicated that the local frequency of these galantamine-induced jaw movements occurs in the 3-7 Hz frequency range that is characteristic of parkinsonian tremor. In the final experiment, the adenosine A(2A) antagonist MSX-3 significantly attenuated the tremulous jaw movements induced by the 3.0mg/kg dose of galantamine, which is consistent with the hypothesis that co-administration of adenosine A(2A) antagonists may be beneficial in reducing parkinsonian motor impairments induced by anticholinesterase treatment.

Longitudinal evaluation of apparent diffusion coefficient in children with neurofibromatosis type 1.

OBJECTIVES: Foci of T2-prolongation in both supra- and infratentorial brain in neurofibromatosis type-1 (NF1) patients have been called hamartoma-like NF1 lesions (HLL); however, their behavior is not consistent with this definition. Diffusion-weighted imaging has been used to study structure and cellularity of intracranial lesions. We applied this technique to characterize HLL as they change with time in pediatric patients. METHODS: We retrospectively studied 12 children (ages 2-20 years) with proven NF1. Forty lesions were studied longitudinally on multiple exams for a total of 166 measurements. Apparent diffusion coefficients (ADCs) were also obtained from a comparison group comprising 14 normal children (ages 2-16 years). RESULTS: The ADC for the lesions was generally greater than normal brain. Both supra- and infratentorial lesions had increasing ADC with patient age. This is contrasted to our comparison group of normal subjects who showed decreasing ADC with age. Linear regression analysis of infratentorial lesions yielded a slope of 2.70 x 10(-6) mm2 x s(-1) x year(-1) versus a slope of 17.57 x 10(-6) mm2 x s(-1) x year(-1) for supratentorial lesions. CONCLUSIONS: Our observed increase in ADC of lesions in pediatric patients with NF1 suggests increasing water in the extracellular space and/or decreasing cellularity with patient age. This change may be due to increased number or size of myelin vacuoles in NF1 lesions. Supratentorial lesions had a statistically significant increase in the rate of change of ADC compared with infratentorial lesions which may reflect an intrinsic difference in the lesion or similar lesions expressed in different environments.