Evaluation of a New Mordant Based Haematoxylin Dye (Haematoxylin X) for Use in Clinical Pathology.

Recently, St John's Dermatopathology Laboratory and CellPath Ltd have developed a new patented haematoxylin dye (Haematoxylin X) that utilises a chromium-based mordant (Chromium Sulphate). In this study, the performance of this new haematoxylin (Haematoxylin X) was compared against some commonly utilised alum-based haematoxylins (Carazzi's, Harris' and Mayer's) when used as a part of formalin-fixed paraffin embedded (FFPE) tissue, special stains, immunohistochemical counterstaining and frozen section (Mohs procedure) staining procedures. FFPE sections of different tissue types and frozen skin tissues were sectioned and stained with each haematoxylin subtype to allow for a direct comparison of staining quality. The slides were independently evaluated microscopically by two assessors. A combined score was generated to determine the sensitivity (defined as the intensity of haematoxylin staining being too weak or too strong and the colour of the haematoxylin staining not being blue/black) and specificity (defined as the presence of haematoxylin background staining, uneven staining, and staining deposits) for each of the four haematoxylin subtypes. The scoring criteria were based on the UKNEQAS Cellular pathology techniques assessment criteria. In FFPE tissue, the results for specificity identified Harris haematoxylin scoring the highest (91.2%) followed by Haematoxylin X (88.0%) and Mayer's (87.0%). The sensitivity scores again identified Harris haematoxylin as scoring the highest (95.1%) followed by Haematoxylin X (90.0%) and Mayer's (88.0%). In frozen tissue, the results for specificity identified Haematoxylin X as scoring the highest (85.5%) followed by Carazzi's (80.7%) and Harris' (77.4%). The sensitivity scores again identified Haematoxylin X as scoring the highest (86.8%) followed by Carazzi's (82.0%) and Harris' (81.0%). The results achieved with all four haematoxylins showed a high degree of comparability, with Harris' haematoxylin scoring high scores overall compared to the other four when assessing FFPE sections. This may have been due to familiarity with the use of Harris' haematoxylin in-house. There was also evidence of more pronounced staining of extracellular mucin proteins with Haematoxylin X compared to the other alum haematoxylins that were assessed. Haematoxylin X scored highest when used in frozen section staining. In addition, Haematoxylin X has a potential applications for use in IHC and special stains procedures as a counterstain.

Development of a 3D ultrafast laser written near-infrared spectro-interferometer.

Direct ultrafast laser photoinscription of transparent materials is a powerful technique for the development of embedded 3D photonics. This is particularly adaptable for astrophotonic devices when a number of inputs are required. The process relies essentially on volume fabrication of waveguiding structures in flexible 3D designs and refractive index contrast parameters adjustable for specific spectral ranges. This enables 3D geometry and thus avoids in-plane crossings of waveguides that can induce losses and cross talk in multi-telescope beam combiners. The additional novel capability of the technique allows for the fabrication of high aspect ratio nanostructures nonperturbatively sampling the optical field. Combining ultrafast laser micro- and nanoprocessing with engineered beams, we present here results for the development of chip-sized silica glass integrated robust 3D three-telescope beam combiners in the near-IR range, as well as embedded diffraction gratings, for phase closure analysis and spectro-interferometry applications in astronomy.

Waveguide scattering antennas made by direct laser writing in bulk glass for spectrometry applications in the short-wave IR.

A buried straight waveguide perturbed periodically by six antennas composed of submicronic cylinder voids is entirely fabricated using ultrafast laser photoinscription. The light scattered from each antenna is oriented vertically and is detected by a short-wave IR camera bonded to the surface of the glass with no relay optics. The response of each antenna is analyzed using a wavelength tunable laser source and compared to simulated responses verifying the behavior of the antenna. These results show the good potential of the direct laser writing technique to realize monolithic embedded detectors by combining complex optical functions within a 3D design. A wavelength meter application with a spectral resolution of 150 pm is proposed to demonstrate this combination.

Fibrosis assessment in patients with nonalcoholic fatty liver disease: Adherence to proposed algorithms and barriers to complying with them.

INTRODUCTION AND AIMS: Fibrosis staging in patients with nonalcoholic fatty liver disease (NAFLD) is carried out through the application of stepwise algorithms but there is little real-world data on their use. Our aim was to calculate the number of patients with NAFLD and indeterminate or high risk for fibrosis, assessed through noninvasive scores, that consequently underwent further staging evaluation. MATERIALS AND METHODS: A cross-sectional multicenter cohort study was conducted on patients with NAFLD evaluated by hepatologists within the time frame of June 1 and July 31, 2018. The FIB-4 and NAFLD fibrosis scores were calculated in all the patients, and if at least one of the scores suggested indeterminate or high risk for fibrosis, we believed the patient should have undergone additional fibrosis staging assessment. RESULTS: The study included 238 patients. The median time interval from NAFLD diagnosis and inclusion in the analysis was 12.2 months (IQR 3.0-36.5). A total of 128 (54%) patients had at least one noninvasive score that suggested indeterminate or high risk for fibrosis but studies to confirm the fibrosis grade (elastography, biopsy, etc.) were performed on only 72 (56%). The main barriers encountered by the physicians for applying the staging algorithms were related to health insurance coverage and imaging study costs. CONCLUSIONS: A high percentage of patients with NAFLD were at indeterminate or high risk for fibrosis, according to noninvasive scores, but additional studies were carried out on only half of them, showing low adherence to current recommendations.

Traumatic and non-traumatic spinal cord injury: Demographic characteristics, neurological and functional outcomes. A 7-year single centre experience.

Objective: The aim of this study is to evaluate demographic and clinical characteristics of a population affected by traumatic and non-traumatic spinal cord injury (SCI) and to analyze functional outcomes after rehabilitation. Methods: This study involved 112 SCI patients (75 male and 37 female) admitted at the Neurorehabilitation Unit of the University Hospital of Messina. The neurological outcomes were evaluated according to the American Spinal Injury Association Impairment Scale (AIS) and by using length of stay, Functional Independence Measure (FIM) and Barthel Index (BI). Results: NT-SCI patients were significantly older, numerous (75,89%) and affected by greater lesions when admitted, than T-SCI ones. Most of lesions were incomplete (93%) and associated with paraplegia (71%). FIM and BI outcomes are similar in both groups, even if T-SCI patients showed greater improvement when discharged. No significant differences were found in the length of stay. The most common complication in non-traumatic SCI group was urinary tract infection and this was observed in 25 patients (29,41%). Linear regression models explained 26% of the variance of LOS and 38% of the variance of functional outcome. Functional status on admission was the strongest determinant of LOS and completeness of the lesion was the strongest determinant of functional outcome. Etiology (traumatic versus non-traumatic) was a weak independent determinant of LOS but was not an independent determinant of functional outcome. Conclusion: SCI patient's rehabilitation should be carried out by taking into account etiology of the injury. It is important to consider this information while developing the targets and planning of the rehabilitation program. In particular, older age negatively influence the degree of disability on admission and the entity of functional recovery in both populations. Non-traumatic lesions could have minor benefits after rehabilitation therapy if compared with traumatic ones.

Fibrosis assessment in patients with nonalcoholic fatty liver disease: Adherence to proposed algorithms and barriers to complying with them. / Evaluación de fibrosis en pacientes con enfermedad por hígado graso no alcohólico: adherencia a los algoritmos propuestos y barreras para cumplir con ellos.

INTRODUCTION AND AIMS: Fibrosis staging in patients with nonalcoholic fatty liver disease (NAFLD) is carried out through the application of stepwise algorithms but there is little real-world data on their use. Our aim was to calculate the number of patients with NAFLD and indeterminate or high risk for fibrosis, assessed through noninvasive scores, that consequently underwent further staging evaluation. MATERIALS AND METHODS: A cross-sectional multicenter cohort study was conducted on patients with NAFLD evaluated by hepatologists within the time frame of June 1 and July 31, 2018. The FIB-4 and NAFLD fibrosis scores were calculated in all the patients, and if at least one of the scores suggested indeterminate or high risk for fibrosis, we believed the patient should have undergone additional fibrosis staging assessment. RESULTS: The study included 238 patients. The median time interval from NAFLD diagnosis and inclusion in the analysis was 12.2months (IQR 3.0-36.5). A total of 128 (54%) patients had at least one noninvasive score that suggested indeterminate or high risk for fibrosis but studies to confirm the fibrosis grade (elastography, biopsy, etc.) were performed on only 72 (56%). The main barriers encountered by the physicians for applying the staging algorithms were related to health insurance coverage and imaging study costs. CONCLUSIONS: A high percentage of patients with NAFLD were at indeterminate or high risk for fibrosis, according to noninvasive scores, but additional studies were carried out on only half of them, showing low adherence to current recommendations.

Semi-automated standardisation of melanin bleaching procedures of heavily pigmented melanocytic lesions for immunohistochemical analysis on an automated platform.

Background: The diagnosis of heavily pigmented melanocytic lesions is problematic. This is often compounded by lack of visibility of nuclear detail of tumour cells due to physical masking by melanin pigment. Similarly, there can be colour merging of chromogenic final reaction products with melanin, making an evidence of antigenic localisation problematic. There are a number of melanin bleaching techniques available for immunohistochemical assessments.Material and methods: All methods to date have involved the bleaching of melanin as a manually performed primary step before loading subsequently bleached slides onto automated immunohistochemical platforms. Here we define a semi-automated bleaching procedure that allows full integration on one of the most widely employed automated IHC staining platforms (Roche Ventana BenchMark Ultra). The bleaching protocol was defined on the BenchMark Ultra and involved the assessment of 24 histological cases of heavily pigmented malignant melanoma lesions (13 cutaneous and 11 metastatic) routinely fixed processed and paraffin wax embedded.Results: Completion of the bleaching was assessed on H&E preparations performed following the semi-automated bleaching step and employing the Roche Ventana BenchMark Ultra machine for 60 min at 42°C. Complete immunohistochemical staining was achieved on the automated platform within 5-6 h including the bleaching step. Results were consistent across all tissue evaluated.Discussion: This data provides evidence that the hydrogen peroxide bleaching procedure can be adapted for integration on one of the most widely employed automated IHC staining platforms and as a result, improve the efficiency and reproducibility of the technique.

Efficient point-by-point Bragg gratings fabricated in embedded laser-written silica waveguides using ultrafast Bessel beams.

We demonstrate highly efficient Bragg gratings written point-by-point by sequential single-pulse ultrashort Bessel laser beams in laser photoinscribed single-mode waveguides in bulk fused silica. The use of chirped non-diffractive Bessel beams determines a strong Bragg resonance in a weak-to-strong transitional regime, augmenting to a record value of 40 dB/cm at 1550 nm in the third order. The Bessel-induced refractive index modulation is negative and localized to sub-micrometer (200 nm) transverse scales. The strong light confinement in Bessel beams ensuring uniform one-dimensional void conditions thus allows for enhanced precision in the Bragg grating waveguide design. We demonstrate flexible fabrication of multiplexed waveguide gratings for multiple and tunable spectral resonances.

Access to direct-acting antivirals for the treatment of hepatitis C in a country with limited resources. / Evaluación del acceso a antivirales para el tratamiento de la hepatitis C en un país con recursos limitados.

AIMS: To estimate the number of patients that have access to treatment of hepatitis C with direct-acting antivirals in Argentina and evaluate the factors associated with the lack of access. MATERIALS AND METHODS: A cross-sectional cohort study was conducted that included all the consecutive prescriptions of direct-acting antivirals issued at health centers that participated in the ECHOTM telemedicine project directed by the Hospital Italiano de Buenos Aires, within the time frame of January 2016 and February 2017. RESULTS: A total of 143 treatment prescriptions were included and overall access was 70% (95% CI 62-77%). The only independent factor associated with a lack of treatment access was coverage by a public healthcare system (OR 4.98 [95% CI 2.05- 12.09]). CONCLUSION: Patients with hepatitis C that were covered by a public healthcare system had a 4 times higher chance of not having access to treatment with direct-acting antivirals than patients covered by other healthcare systems (private insurance or the social welfare system).

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.

Near infrared spectro-interferometer using femtosecond laser written GLS embedded waveguides and nano-scatterers.

Guided optics spectrometers can be essentially classified into two main families: based on Fourier transform or dispersion. In the first case, an interferogram generated inside an optical waveguide and containing the spectral information is sampled using spatially distributed nanodetectors. These scatter quasi-non-perturbingly light into the detector that is in contact with the waveguide, helping to reconstruct the stationary wave. A dedicated FFT processing is needed in order to recover the spectrum with high resolution but limited spectral range. Another way is to directly disperse the different wavelengths to different pixels, either introducing differential optical path in the same propagation plane (multiple Mach-Zehnder interferometers or Arrayed Waveguides Gratings), or using a periodic structure to perpendicularly extract the optical signal confined in a waveguide (photonic crystals or surface gratings), and by means of a relay optics, generate the spectrum on the Fourier plane of the lens, where the detector is placed. Following this second approach, we present a laser-fabricated high-resolution compact dispersive spectro-interferometer (R>2500, 30nm spectral range at λ = 1560nm), using four parallel waveguides that can provide up to three non-redundant interferometric combinations. The device is based on guided optics technology embedded in bulk optical glass. Ultrafast laser photoinscription with 3D laser index engineering in bulk chalcogenide Gallium Lanthanium Sulfide glass is utilized to fabricate large mode area waveguides in an evanescently-coupled hexagonal multicore array configuration, followed by subsequent realization of nanoscaled scattering centers via one dimensional nanovoids across the waveguide, written in a non-diffractive Bessel configuration. A simple relay optics, with limited optical aberrations, reimages the diffracted signal on the focal plane array, leading to a robust, easy to align instrument.

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.

Low-power-threshold photonic saturable absorber in nonlinear chalcogenide glass.

We experimentally demonstrate controllable nonlinear modulation of optical guiding in ultrafast laser-written evanescently coupled waveguide arrays in bulk gallium lanthanum sulfide chalcogenide glass. The intensity-dependent response is validated by simulating light propagation in waveguide arrays with instantaneous Kerr nonlinearity using a discrete-continuous spatiotemporal unidirectional Maxwell equation model. The intensity-driven modulation of transmission in multicore structures acts as a potential saturable absorber at kilowatt threshold levels.

Large-mode-area infrared guiding in ultrafast laser written waveguides in sulfur-based chalcogenide glasses.

Current demands in astrophotonics impose advancing optical functions in infrared domains within embedded refractive index designs. We demonstrate concepts for large-mode-area guiding in ultrafast laser photowritten waveguides in bulk Sulfur-based chalcogenide glasses. If positive index contrasts are weak in As2S3, Ge doping increases the matrix rigidity and allows for high contrast (10(-3)) positive refractive index changes. Guiding with variable mode diameter and large-mode-area light transport is demonstrated up to 10 µm spectral domain using transverse slit-shaped and evanescently-coupled multicore traces.

Large mode area waveguides with polarization functions by volume ultrafast laser photoinscription of fused silica.

We present optical designs allowing large mode area light guiding by ultrafast laser photoinscription of bulk fused silica. If usual concepts are based on large core and depressed cladding, evanescently coupled multicore waveguides with coherent mode superposition can be effective solutions, where the introduction of nanostructured defects determines additional polarization functions.

Association of low-penetrance alleles with male breast cancer risk and clinicopathological characteristics: results from a multicenter study in Italy.

It is well-known that male breast cancer (MBC) susceptibility is mainly due to high-penetrance BRCA1/2 mutations. Here, we investigated whether common low-penetrance breast cancer (BC) susceptibility alleles may influence MBC risk in Italian population and whether variant alleles may be associated with specific clinicopathological features of MBCs. In the frame of the Italian Multicenter Study on MBC, we genotyped 413 MBCs and 745 age-matched male controls at 9 SNPs annotating known BC susceptibility loci. By multivariate logistic regression models, we found a significant increased MBC risk for 3 SNPs, in particular, with codominant models, for rs2046210/ESR1 (OR = 1.71; 95 % CI: 1.43-2.05; p = 0.0001), rs3803662/TOX3 (OR = 1.59; 95 % CI: 1.32-1.92; p = 0.0001), and rs2981582/FGFR2 (OR = 1.26; 95 % CI: 1.05-1.50; p = 0.013). Furthermore, we showed that the prevalence of the risk genotypes of ESR1 tended to be higher in ER- tumors (p = 0.062). In a case-case multivariate analysis, a statistically significant association between ESR1 and ER- tumors was found (OR = 1.88; 95 % CI: 1.03-3.49; p = 0.039). Overall, our data, based on a large and well-characterized MBC series, support the hypothesis that common low-penetrance BC susceptibility alleles play a role in MBC susceptibility and, interestingly, indicate that ESR1 is associated with a distinct tumor subtype defined by ER-negative status.

A multicenter phase II randomized trial of docetaxel/gemcitabine versus docetaxel/capecitabine as first-line treatment for advanced breast cancer: a Gruppo Oncologico Italia Meridionale study.

OBJECTIVE: To evaluate two docetaxel-based regimens as first-line treatment in advanced breast cancer patients. METHODS: Patients were randomly assigned to docetaxel/gemcitabine (arm A: docetaxel 75 mg/m(2) on day 1, gemcitabine 1,000 mg/m(2) on days 1 and 8) or docetaxel/capecitabine (arm B: docetaxel 75 mg/m(2) on day 1, capecitabine 1,250 mg/m(2) twice daily on days 1-14); both chemotherapy regimens were repeated every 21 days. The primary objective of the study was to evaluate the response rate. RESULTS: Seventy-two patients were enrolled (36 each in arms A and B). Responses according to intention-to-treat analysis were as follows: arm A, 41.7% [95% confidence interval (CI) 25.6-57.8]; arm B, 38.9% (95% CI 23-54.8). Median progression-free survival was 10.9 months (95% CI 8.1-13.7) in arm A and 10 months (95% CI 8.8-11.2) in arm B. Overall survival was 26 months (95% CI 22.0-30.0) in arm A and 28 months (95% CI 23.4-32.6) in arm B. Both treatments were well tolerated; myelosuppression was the dose-limiting toxicity, with grade 3-4 neutropenia in 13.8 and 19.4% of the patients in arms A and B, respectively. No relevant differences in other toxicities were observed in the two arms, except for diarrhea (13.9%) and hand-foot syndrome (11.1%), which occurred only in arm B. CONCLUSIONS: Both regimens were active and well tolerated in advanced breast cancer.

Diet assessment and the screening for osteoporosis: a survey in a healthy menopause population.

AIM: The aim of this study was to investigate about diet assessment in the screening for osteoporosis. METHODS: In 1,441 women (group A, mean age 53.5 years, mean time from menopause 3.92(+/-4.33)] the number of daily/weekly servings/cups of different foods has been collected. In 71 women (group B, mean age 59 years, mean time from menopause 6 years) diet was assessed through a detailed questionnaire. Demographic and clinical data and dietary habits were related to bone mineral density (BMD) values, using Student t-test and the chi2 test. RESULTS: Group A: borderline/reduced BMD 40%, the percentage increasing with age and the duration of menopause (P<0.001). BMI within normal range 56.28%. BMD values were positively associated with Body Mass Index (BMI, P<0.001), education level (P<0.005), job (P<0.01), oral contraceptives use (P<0.001), negatively with family history of osteoporosis (P<0.05). BMD values showed a positive relation with the number of eggs per week (P<0.05), positive not significant trends were observed with dairy products, vegetables, red and white meat intake. A low/reduced BMD was related to low calcium food intake in less than 50% of the women. Group B: borderline/reduced BMD 55%; BMD showed positive trends, not significant, with daily calcium intake, BMI, physical exercise. Estimated calcium intake less than 700 mg/day was found in 33% of the sample. CONCLUSION: Diet assessment is not useful in the screening for osteoporosis. In women with borderline or reduced BMD, diet assessment helps to detect those at low calcium intake who need calcium supplementation, alone or in combination with other treatments.

Conical forward THz emission from femtosecond-laser-beam filamentation in air.

We attribute a strong forward directed THz emission from femtosecond laser filaments in air to a transition-Cherenkov emission from the plasma space charge moving behind the ionization front at light velocity. Distant targets can be easily irradiated by this new source of THz radiation.

Foam sclerotherapy with Fibrovein (STD) for the treatment of hemorrhoids, using a flexible endoscope.

AIM: This case study was designed to evaluate foam sclerotherapy of hemorrhoidal disease using a flexible endoscope. METHODS: We treated 250 patients of grade II-IV hemorrhoids with Sotradecol foam. To obtain sclerosing foam, we used a special foam kit, which produced a stable and standardized sclerosing foam. The quantity of foam injected into the hemorrhoids was 1 to 2 cc with a total of 6 cc for each session. Variables to assess efficacy of treatment were: pain, bleeding and prolapse. RESULTS: Generally, bleeding and hemorrhoidal prolapse were resolved with at most two sclerotherapy sessions. However, pain usually had disappeared after the first session. No complications such as mucosal erosions at the foam injection site, formation of abscesses, bacteraemia, or fistulas occurred. CONCLUSION: The advantages of sclerosing foam are its adhesiveness and compactness, its greater volume, its increased spasm generation, and the greater sclerosing power. Sclero-therapy with foam makes it possible to reach optimal results with small amounts of sclerosing agent and fewer risks of complications.