Standard Histopathological Analysis Research Articles

Histopathological study of sheep lung roaming in dump zones

Respiratory system disease is common in sheep, which is frequently difficult to detectable in live animals until it becomes serious and complicated. This study was aimed to describe sheep lung lesions in Mosul city, which roam in dump zones and have been slaughtered at butchers’ shops, fifty male and female sheep 1-5 years were used at the current work from March to December 2021; affected Samples were collected and prepared for standard histopathological analysis. The special stain was also used for carbohydrates demonstrated inside and around lung lesions. The results of the current study showed the presence of gross and histological finding changes, the gross lesion presented by inflammation of the upper right lope, congestion, and bleeding on all parts of the lope in addition to different sizes of the nodular lesion. In contrast, the pneumonic histological finding was 12.1%, circulatory disturbance 3.5%, parasitic infections 10.0%, disturbance of growth 7.1%, Necrotic area surround by swirling macrophage 2.0% in addition to the deposition of calcium salt and hemosiderin pigmentation. Organizing Pneumonia (OP) and Masson’s bodies were one of the most detected lesions in the roaming sheep, it was scored by Masson Trichrome from mild, moderate, and severe fibrosis. All lesions were categorized according to the type of lesion, grade, and kind of it. From the current work, we demonstrated that pneumonic disorder represented a considerable and serious problem in the animal livestock industry and leads to economic loss in milk, wool, and sheep meat production. Additional studies are recommended to determine the etiological agent of sheep lung lesions whether it is a fungal, bacterial, or viral agent.

Label-free multimodal nonlinear optical microscopy reveals features of bone composition in pathophysiological conditions.

Bone tissue features a complex microarchitecture and biomolecular composition, which determine biomechanical properties. In addition to state-of-the-art technologies, innovative optical approaches allowing the characterization of the bone in native, label-free conditions can provide new, multi-level insight into this inherently challenging tissue. Here, we exploited multimodal nonlinear optical (NLO) microscopy, including co-registered stimulated Raman scattering, two-photon excited fluorescence, and second-harmonic generation, to image entire vertebrae of murine spine sections. The quantitative nature of these nonlinear interactions allowed us to extract accurate biochemical, morphological, and topological information on the bone tissue and to highlight differences between normal and pathologic samples. Indeed, in a murine model showing bone loss, we observed increased collagen and lipid content as compared to the wild type, along with a decreased craniocaudal alignment of bone collagen fibres. We propose that NLO microscopy can be implemented in standard histopathological analysis of bone in preclinical studies, with the ambitious future perspective to introduce this technique in the clinical practice for the analysis of larger tissue sections.

A System-based Approach for the Evaluation of Electromechanical Properties of Brain Tumors.

We have developed a semi-automated system integrated with MEMS-based electromechanical sensors to characterize human brain tumors. The electrical impedance and elastic moduli of three types of brain tumors and six normal brain regions were evaluated using the system. The impedance and elastic modulus of glioma was found to be significantly lower than the normal region. It was also observed that the white matter tissues had higher impedance and elastic moduli compared with the grey matter of the same neuroanatomic location. There were observable differences in the electromechanical behavior of gliomas, which originate from glial cells to that of schwannoma and meningioma of different cellular origins. Clinical Relevance- The observations suggest that simultaneous electromechanical characterization of brain tumors can serve as an effective tool for tumor delineation. The developed tool can be used alongside gold standard histopathological analysis to better understand human brain tumors.

Phosphorylated TDP-43 aggregates in peripheral motor nerves of patients with amyotrophic lateral sclerosis.

Phosphorylated TDP-43 (pTDP-43) aggregates in the cytoplasm of motor neurons and neuroglia in the brain are one of the pathological hallmarks of amyotrophic lateral sclerosis. Although the axons exceed the total volume of motor neuron soma by several orders of magnitude, systematic studies investigating the presence and distribution of pTDP-43 aggregates within motor nerves are still lacking. The aim of this study is to define the TDP-43/pTDP-43 pathology in diagnostic motor nerve biopsies performed on a large cohort of patients presenting with a lower motor neuron syndrome and to assess whether this might be a discriminating tissue biomarker for amyotrophic lateral sclerosis and non-amyotrophic lateral sclerosis cases.We retrospectively evaluated 102 lower motor neuron syndrome patients referred to our centre for a diagnostic motor nerve biopsy. Histopathological criteria of motor neuron disease and motor neuropathy were applied by two independent evaluators, who were blind to clinical data. TDP-43 and pTDP-43 were evaluated by immunohistochemistry, and results compared to final clinical diagnosis.We detected significant differences between amyotrophic lateral sclerosis and non-amyotrophic lateral sclerosis cases in pTDP-43 expression in myelinated fibres: axonal accumulation was detected in 98.2% of patients with amyotrophic lateral sclerosis versus 30.4% of non-amyotrophic lateral sclerosis samples (P < 0.0001), while concomitant positive staining in Schwan cell cytoplasm was found in 70.2% of patients with amyotrophic lateral sclerosis versus 17.4% of patients who did not have amyotrophic lateral sclerosis (P < 0.001). Importantly, we were also able to detect pTDP-43 aggregates in amyotrophic lateral sclerosis cases displaying normal features at standard histopathological analysis.Our findings demonstrated that a specific pTDP-43 signature is present in the peripheral nervous system of patients with amyotrophic lateral sclerosis, and could be exploited as a specific, accessible tissue biomarker. The detection of pTDP-43 aggregates within motor nerves of living patients with amyotrophic lateral sclerosis, occurring before axonal degeneration, suggests that this is an early event that may contribute to amyotrophic lateral sclerosis pathogenesis.

Hormone treatment and UVB exposure influences on female mice regarding skin physiological parameters, biochemical parameters and organ histology

Females require at a certain period of life the administration or supplementation of specific hormones (estrogen, progesterone), for various needs, such as: prevention of unwanted pregnancies, decreased menstrual bleeding, dysmenorrhea and pelvic pain in endometriosis, alleviation of symptoms associated with menopause, regulation of certain skin processes related to acne or aging and others. Also, hormones could act as oncogenes being known eloquent examples of estrogens labeled both as promoters of cell specific alteration or as mutagenic agents. The use of hormones and exposure to solar radiation is expected to cause a number of adverse changes to the body, especially due to their association with malignant processes. The current study was purported as a basis for understanding certain processes that occur with the administration of hormones and exposure to ultraviolet B (UVB) radiation. The animal model was made on healthy adult female BALB/c mice, which were separated into groups and treated with Ethinylestradiol (EES), Levonorgestrel (LNG) and their combination in the presence of UVB radiation. Changes in skin physiological parameters were analyzed by non-invasive methods, biochemical parameters related to changes in blood circulating system were evaluated by standard methods and histopathological analysis was conducted to point out the changes at the level of the internal body. Measurement of skin parameters such as erythema, melanin, skin hydration, has highlighted some changes in hormone-treated and exposed to UVB radiation groups which were significant only in the case of erythema. Biochemical parameters showed variations in terms of liver enzymes in groups treated with active substances. Histologically, aspects of internal organs revealed significant changes in the group treated with EES and LNG and exposed to UVB radiation.

Experimental validation of depolarizing Mueller matrix model via ex vivo colon samples

Experimental validation of a previously reported depolarizing Mueller matrix model is crucial to understanding the light-tissue interactions and the morphological alterations originating in malignant tissue zones in terms of their optical properties. This manuscript is a continuation of a previously reported case study, where a theoretical model was introduced with an additional Monte Carlo simulation. The main aim of this study is to empirically validate both theory and modelling. Once extracted from an experimental Mueller matrix via symmetric decomposition, the polarimetric quantities were compared with the corresponding parameters previously determined from polarized Monte Carlo simulations. Our results were obtained from an ex vivo colon sample and indicate a potential capability to provide supplementary polarimetric data to physicians and the gold standard histopathology analysis.

Simultaneous Characterization of Implant Wear and Tribocorrosion Debris within its Corresponding Tissue Response Using Infrared Chemical Imaging

Biotribology is one of the key branches in the field of artificial joint development. Wear and corrosion are among fundamental processes which cause material loss in a joint biotribological system; the characteristics of wear and corrosion debris are central to determining the in vivo bioreactivity. Much effort has been made elucidating the debris-induced tissue responses. However, due to the complexity of the biological environment of the artificial joint, as well as a lack of effective imaging tools, there is still very little understanding of the size, composition, and concentration of the particles needed to trigger adverse local tissue reactions, including periprosthetic osteolysis. Fourier transform infrared spectroscopic imaging (FTIR-I) provides fast biochemical composition analysis in the direct context of underlying physiological conditions with micron-level spatial resolution, and minimal additional sample preparation in conjunction with the standard histopathological analysis workflow. In this study, we have demonstrated that FTIR-I can be utilized to accurately identify fine polyethylene debris accumulation in macrophages that is not achievable using conventional or polarized light microscope with histological staining. Further, a major tribocorrosion product, chromium phosphate, can be characterized within its histological milieu, while simultaneously identifying the involved immune cell such as macrophages and lymphocytes. In addition, we have shown the different spectral features of particle-laden macrophages through image clustering analysis. The presence of particle composition variance inside macrophages could shed light on debris evolution after detachment from the implant surface. The success of applying FTIR-I in the characterization of prosthetic debris within their biological context may very well open a new avenue of research in the orthopedics community.

Coenzyme Q10 nullified khat-induced hepatotoxicity, nephrotoxicity and inflammation in a mouse model

Ethnopharmacological relevanceThe consumption of khat (Catha Edulis, Forsk) is on the rise despite the much publicized associated deleterious health effects. How chemicals present in khat, affect various physiological and biochemical processes requires further scrutiny. A clear understanding of these processes will provide an avenue for countering khat-driven negative effects using appropriate pharmacological and/or nutritional interventions. Aim of the studyThe current study investigated the effect of khat on vital physiological and biochemical processes such as oxidative stress, inflammation and immune responses and the role of Coenzyme-Q10 (CoQ10), a potent antioxidant and anti-inflammatory, in modulating any negative effects due to khat exposure. MethodologyThree (3) weeks old forty (40) Swiss albino mice were randomly assigned into four treatment groups (n = 10). The first group was the control that was not administered with khat or CoQ10. The second group received 200 mg/kg body weight (b/w) of CoQ10, while the third group received 1500 mg/kg b/w of khat extract and finally the forth group was co-treated with 200 mg/kg b/w of CoQ10 and 1500 mg/kg b/w of khat extract. The experiment was conducted for 90 days after which samples were collected for physiological and biochemical analyses. ResultsThe effects of khat and CoQ10 on the weights of brain, liver, kidney and spleen was determined. Administration of khat decreased the levels of RBCs and its subtypes (MCV, MCH, RDW-SD and RDW-CV), a clear indicator of khat-induced normochromic microcytic anemia. White blood cells (lymphocytes, monocytes, neutrophils and eosinophil) which are vital in responding to infections were markedly elevated by khat. Moreover, these results provide evidence for khat-induced liver and kidney injury as shown by increased biomarkers; AST, ALT, GGT and creatinine respectively. Standard histopathological analysis confirmed this finding for khat-driven liver and kidney injury. Further studies showed evidence for khat-induced inflammation and oxidative stress as depicted by increased levels of the pro-inflammatory cytokine TNF-alpha and elevation of GSH in the brain, liver and spleen. Remarkably, this is the first study to demonstrate the potential of CoQ10 in ameliorating khat-induced negative effects as outlined. CoQ10 supplementation restored the khat-induced reduction in RBC subtypes, and was protective against liver and kidney injury as shown by the appropriate biomarkers and standard histopathology analysis. The other significant finding was the CoQ10-driven normalization of GSH and TNF-α levels, indicating a protective effect from khat-driven oxidative stress and inflammation respectively. ConclusionFrom this study, we conclude that CoQ10 may be useful in nullifying khat-driven deleterious events among chronic khat users.

Intraoperative Detection of Pediatric Brain Tumor Margins Using Raman Spectroscopy.

Abstract INTRODUCTION Surgical resection is a mainstay of treatment in patients with brain tumors both for tissue diagnosis and for tumor debulking. While maximal resection of tumors is desired, neurosurgeons can be limited by the challenge of differentiating normal brain from tumor using only microscopic visualization and tactile feedback. Additionally, intraoperative decision-making regarding how aggressively to pursue a gross total resection frequently relies on pathologic preliminary diagnosis using frozen sections which are both time consuming and fallible. Here, we investigate the potential for Raman spectroscopy (RS) to rapidly detect pediatric brain tumor margins and classify brain tissue samples equivalent to histopathology. METHODS Using a first-of-its-kind rapid acquisition RS device we intraoperatively imaged fresh ex vivo pediatric brain tissue samples (2-3 mm × 2-3 mm × 2-3 mm) at the Lucille Packard Children's Hospital. All imaged samples received standard final histopathological analysis, as RS is a nondestructive imaging technique. We curated a labeled dataset of 575 + unique Raman spectra gathered from 160 + brain samples resulting from 23 pediatric patients who underwent brain tissue resection as part of tumor debulking or epilepsy surgery (normal controls). RESULTS To our knowledge we have created the largest labeled Raman spectra dataset of pediatric brain tumors. We are developing an end-to-end machine learning model that can predict final histopathology diagnosis within minutes from Raman spectral data. Our preliminary principle component analyses suggest that RS can be used to classify various brain tumors similar to “frozen” histopathology and can differentiate normal from malignant brain tissue in the context of low-grade glioma resections. CONCLUSION Our work suggests that machine learning approaches can be used to harness the material identification properties of RS for classifying brain tumors and detecting their margins.

HIF1α and SOX2 expression in cervical squamous cell carcinoma

Background/Aim. Hypoxia is one of the major changes that occurs in the tumor microenvironment. It has been observed that there are pluripotent cancer cells in the cancer cell population that affect tumor growth and their resistance to therapy. The aim of this study was to examine the expression of hypoxia-inducible factor-1 alpha (HIF-1?), endogenous marker of hypoxia, and SOX2, marker of the pluripotent stem cells existing in the normal adult tissues, in the cervical squamous cell carcinoma (SCC). Methods. The study was conducted in 90 women with invasive cervical SCC, divided into two groups ? 60 women in the Group A, with FIGO IB1 &lt; 20 mm tumors (no metastases in the lymph nodes), and 30 women in the group B with tumors FIGO I?II (positive lymph nodes). The basic clinical parameters were determined by standard histopathological analysis, and the expression of HIF-1? and SOX2 by immunohistochemical examination. Results. There was a significant difference between the groups A and B, in the expression of HIF-1? (p = 0.024), but not in the expression of SOX2 (p = 0.566). Expression of HIF-1? was significantly higher in the group with lymph node metastases and invasion of lymphovascular spaces (p &lt;0.001) but not associated with tumor size (p = 0.291) or lymphocytic stromal response (p = 0.940). The tumor grade significantly influenced the expression of HIF-1? (p = 0.013). The expression of SOX2 did not significantly correlate with any of the established clinical tumor parameters. Conclusion. A significant difference in the expression of HIF-1 ? between the group with and that without metastases in lymph nodes in invasive cervical SCC could distinguish HIF-1? as a parameter of poor prognosis of the disease. The prognostic significance of SOX2 as well as a significant correlation between expression of HIF-1? and SOX2 were not established.

Smartphone confocal microscopy for imaging cellular structures in human skin in vivo.

We report development of a low-cost smartphone confocal microscope and its first demonstration of in vivo human skin imaging. The smartphone confocal microscope uses a slit aperture and diffraction grating to conduct two-dimensional confocal imaging without using any beam scanning devices. Lateral and axial resolutions of the smartphone confocal microscope were measured as 2 and 5 µm, respectively. In vivo confocal images of human skin revealed characteristic cellular structures, including spinous and basal keratinocytes and papillary dermis. Results suggest that the smartphone confocal microscope has a potential to examine cellular details in vivo and may help disease diagnosis in resource-poor settings, where conducting standard histopathologic analysis is challenging.

Abstract P2-12-05: Real-time, intraoperative detection of residual breast cancer in lumpectomy cavity margins using the LUM imaging system: Results of a feasibility study

Abstract Background: Obtaining tumor-free margins is critical for local control in breast conserving surgery. Currently, 20-40% of lumpectomy patients have positive margins that require surgical re-excision. We assessed the LUM Imaging System for real-time, intraoperative detection of residual tumor in breast cancer patients. The LUM System has the particular advantage of assessing in vivo lumpectomy cavity walls rather than excised specimens, to enable more accurate excision of residual tumor. Methods: Lumpectomy cavity walls of patients undergoing lumpectomy for invasive breast cancer or ductal carcinoma in situ (DCIS), were assessed intraoperatively using the LUM Imaging System (Lumicell Inc., Wellesley MA). LUM015, a cathepsin-activatable fluorescent agent, was given IV 4±2 hrs prior to surgery. Areas of fluorescence generated at potential sites of residual tumor in lumpectomy cavities were evaluated with a sterile hand-held device, displayed on a monitor, excised and correlated with histopathology. Results: In vivo lumpectomy cavities were imaged with the LUM Imaging System in 60 breast cancer patients. 5 were imaged without dye. 55 received LUM015 dye preoperatively and were scanned intraoperatively. Median age was 60 years (range 44-79). Mean tumor size was 1.2cm (0.06-3.5cm) with 71% invasive cancers, 29% DCIS. The test set included 569 cavity margin surfaces assessed intraoperatively and excised. Image acquisition for each margin took approximately 1 second. The LUM Imaging System showed 100% sensitivity and 73% specificity for detection of tumor &amp;lt;2mm from the margin. Invasive ductal cancer (IDC), invasive lobular cancer (ILC) and areas of DCIS 1mm in size could be identified. 8 patients had positive margins on standard histopathology analysis (Table). The LUM System correctly identified all positive margins identified by standard histopathology and correctly predicted negative re-excisions in 2 of 8 patients. There were no serious adverse events. 1 patient had extravasation of LUM015 at her injection site with temporary blue skin staining but no other complication. Conclusions: The LUM Imaging System allows real-time identification of residual tumor in the lumpectomy cavity of breast cancer patients. No sites of residual tumor were missed. Additional studies are underway to optimize this approach for reducing positive margins and second surgeries in breast cancer patients. Table: Margin results in 8 patients with positive margins on initial lumpectomy specimenPositive lumpectomy margin histopathologyLUM cavity wall result (+/- for tumor)Tumor found at re-excisionDCIS++DCIS+-DCIS++IDC++ (Mastectomy)ILC++ (Mastectomy)DCIS+-IDC--DCIS-- Citation Format: Lanahan CR, Gadd MA, Specht MC, Ferrer J, Tang R, Rai U, Merrill AL, Biernacka A, Brachtel E, Smith BL. Real-time, intraoperative detection of residual breast cancer in lumpectomy cavity margins using the LUM imaging system: Results of a feasibility study [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P2-12-05.

Detection of Placental Chromosomal Aberrations in Early Spontaneous Abortionsin Correlation with the Histologic Findings

Abstract Using a variety of molecular techniques, it has been established that loss of pregnancy occurs in one to two thirds of all fertilized embryos in the first trimester. In about 50% of the cases, chromosomal abnormalities are the cause of early spontaneous abortion. Several histological characteristics of the placenta, such as presence of villous stromal cavitations, fetal erythrocytes, umbilical cord, fetal tissue, etc. are suggested as predictive factors for aneuploidy. Two hundred and thirty one cases were analyzed in this prospective study, 50 cases were control artificial abortions and 181 cases were early spontaneous abortionsanalyzed in the period from May 2012 to December 2014. Standard histopathological analysis and molecular techniques based on polymerase chain reaction were used to analyze the samples. Usingmolecular techniques, aneuploidy was detected in 53.1% of the samples. The most frequently detected aneuploidy was trisomy 16, followed by trisomy 22, 21, 14 and 18. The molecular analysis also enabled distinction of maternal and paternal origin of the alleles. In the histopathological sample analysis, binary logistic regression analysis indicated the presence of trophoblastic proliferation (p=0.008) and the absence of fetal red blood cells (p=0.001) as independent significant factors in the prediction of aneuploidy in early spontaneous abortion. In conclusion, our results show that clinically relevant and accurate diagnosis of early spontaneous abortion which can determine its causecan only be achieved bya controlled process of selection of the material, histo-pathological and molecular analysis, followed bya necessary correlation of these results.

Molecular and histological characteristics of early triploid and partial molar pregnancies

Molar pregnancy has the highest incidence of all gestational trophoblastic diseases. This is a heterogeneous group of diseases, composed of precancerous lesions and gestational trophoblastic tumours. The hydatidiform mole is characterised by varying degrees of proliferation of syncytiotrophoblastic and cytotrophoblastic cells and stromal oedema. Based on established morphological and cytogenetic criteria, molar pregnancy is divided into partial and complete. The risk of persistent trophoblastic disease is higher in complete moles compared with partial moles. The aim of this study was to assess the importance of additional molecular methods as a conjunction to the standard histopathological analysis to accurately determine the type and origin of triploidy and to detect partial molar pregnancy. This study examined a total of 24 cases of triploidy. Apart from the detailed histomorphological analysis, a molecular analysis of the placental tissue and maternal DNA was also performed. Digynic triploidy was found in 15 cases, whereas diandric triploidy was found in nine of the cases. The results showed that due to the histomorphological overlap between partial molar pregnancy and hydropic abortions, concomitant histopathological analysis of the placental tissue and molecular analysis of the placental and maternal DNA can lead to correct diagnosis.

Single punch, double biopsy.

Objective In lethal primary metastatic prostate cancer, biopsy material is often the only accessible cancer tissue. Lack of tissue quantity limited the use of biopsy cores for analyzing higher numbers of molecular markers and standard histopathologic evaluation for clinical diagnosis simultaneously. Recent advances in single cell analytics have paved the way to characterize a tumor in more depth from minute input material such as biopsies. We therefore aimed to develop a biopsy needle, which generates two cores side by side from the same punch: one for standard histopathologic analysis to allow for routine diagnostics and the second one for single cell analytics.Methods On the basis of a conventional punch biopsy needle we have milled two parallel longitudinal rifts into the needles shat which are separated by a 100 µm thick metal sheet. Each rift can harbor a single tissue core.ResultsTwo cores from the same punch were generated reproducibly from a radical prostatectomy specimen and showed congruent results in histopathologic analysis. Both cores yielded equally sufficient material for standard H&E staining and histopathological evaluation.ConclusionOur modified biopsy system will allow for simultaneous acquisition of tissue cores for diagnostic and scientific analysis from solid tumors or metastatic sites.

A Pilot Study of In Vivo Confocal Laser Endomicroscopy of Upper Tract Urothelial Carcinoma.

Tissue diagnosis of upper tract urothelial carcinoma (UTUC) is limited by variance in tumor sampling by standard ureteroscopic biopsy. Optical imaging technologies can potentially improve UTUC diagnosis, surveillance, and endoscopic treatment. We previously demonstrated in vivo optical biopsy of urothelial carcinoma of the bladder using confocal laser endomicroscopy (CLE). In this study, we evaluated a new 0.85-mm imaging probe in the upper urinary tract and demonstrated feasibility and compatibility with standard ureteroscopes to achieve in vivo optical biopsy of UTUC. Fourteen patients scheduled for ureteroscopy of suspected upper tract lesions or surveillance of UTUC were recruited. After intravenous (IV) administration of fluorescein, CLE was performed using a 0.85-mm-diameter imaging probe inserted through the working channel of standard ureteroscopes. Acquired confocal video sequences were reviewed and analyzed. A mosaicing algorithm was used to compile a series of images into a single larger composite image. Processed CLE images were compared with standard histopathologic analysis. Optical biopsy of the UTUC using CLE was effectively achieved during standard ureteroscopy. There were no adverse events related to IV fluorescein administration or image acquisition. Confocal imaging of UTUC showed characteristic features similar to urothelial carcinoma of the bladder, including papillary structure, fibrovascular stalks, and pleomorphism. Lamina propria in normal areas of the renal pelvis and ureter was also identified. We report an initial feasibility of CLE of UTUC. Pending further clinical investigation, CLE may become a useful adjunct to ureteroscopic biopsy, endoscopic ablation, and surveillance of UTUC.

Feasibility of transoral robotic‐assisted high‐resolution microendoscopic imaging of oropharyngeal squamous cell carcinoma

Transoral robotic-assisted oncologic surgery of the head and neck offers promising functional results. Nonetheless, the efficacy of oncologic surgery remains critically dependent on obtaining negative margins. We aimed to integrate a miniaturized high-resolution fiber-optic microendoscope (HRME), which provides real-time histological assessment, with the da Vinci robotic system (Intuitive Surgical, Sunnyvale, CA). Three patients undergoing transoral robotic surgery (TORS) were prospectively enrolled in this study. Optical imaging of the oropharynx was performed intraoperatively with the robotic-assisted HRME. All patients underwent the procedure successfully with no complications. The HRME was successfully integrated with the da Vinci robotic system. Several sites of the oropharynx and associated malignancy were imaged, which correlated with the standard histopathological analysis. Transoral robotic-assisted HRME imaging of the oropharynx is a safe and technically feasible approach, providing a real-time histological assessment and may serve as a valuable aid in oncologic surgery.

Prognostic factors in oral and oropharyngeal cancer based on ultrastructural analysis and DNA methylation of the tumor and surgical margin.

Oral and oropharyngeal cancers are characterized by relatively low 5- year survival rates due to many factors, including local recurrence. The identification of new molecular markers may serve for the estimation of prognosis and thus augment treatment decisions and affect therapy outcome. The aim of this study was to describe the morphological characteristics and the DNA methylation status of the CDKN2A,CDH1, ATM, FHIT and RAR- genes in the central and peripheral part of the tumor and the surgical margin and evaluate their prognostic significance. 53 patients with oral and oropharyngeal cancer were enrolled to the prospective study, and had been primarily treated surgically. Correlations between morphological data, hypermethylation status and clinicopathological data, as well as prognosis, were assessed. Nuclei polymorphism highly correlated with T stage (p < 0.0001), N stage (p < 0.046), and metastases to the lymph nodes pN (p < 0.004 ). Also, the number of cells in irregular mitosis correlated with T stage (p < 0.004), and highly with pN (p < 0.009). The significance of CDKN2A hypermethylation as a good prognostic factor was also established in the Kaplan-Meir test. The ultrastructural analysis showed that none of the examined tumors had homogenous texture and that resection margin specimens clean in HE stained tissue samples frequently contained single tumor cells or few cells in groups surrounded by connective tissue. This indicates the superiority of electron microscopy over standard histopathological analysis. Thus, a combination of such morphological examination with epigenetic parameters described herein could result in the discovery of promising new prognostic markers of the disease.

Cigarette smoke induces molecular responses in respiratory tissues of ApoE−/− mice that are progressively deactivated upon cessation

Cigarette smoking is the primary etiology of chronic obstructive pulmonary disease (COPD) and a risk factor for both lung and cardiovascular (CV) diseases, which are rarely investigated concomitantly. Although smoking cessation shows clear CV risk benefit, lung-related disease risk remains higher in former smokers than in never smokers. We sought to determine the differential molecular responses of murine respiratory tissues to better understand the toxicity pathways involved in smoking-related disease risk and those related to the benefits of smoking cessation. ApoE−/− mice were exposed to mainstream cigarette smoke (CS) or a smoking cessation-mimicking protocol for up to 6 months and transcriptomics analysis of nasal epithelium and lung parenchyma performed. We supported our gene expression profiling approach with standard lung histopathology and bronchoalveolar lavage fluid (BALF) analysis. Many BALF analytes involved in functions ranging from inflammation to cell proliferation and tissue remodeling were found elevated in BALF. Gene expression levels of these molecules were also increased in lung tissue, suggesting that the inflammatory response was the result of local tissue activation and the contribution of recruited inflammatory cells. Gene set enrichment analysis (GSEA) of expression data from murine lungs and nasal epithelium showed distinct activation patterns of inflammation, complement, and xenobiotic metabolism pathways during CS exposure that were deactivated upon smoking cessation. Pathways involved in cell proliferation and tissue remodeling were activated by CS and progressively deactivated upon smoke exposure cessation. Differential CS-mediated responses of pulmonary and nasal tissues reflect common mechanisms but also the varying degrees of epithelial functional specialization and exposure along the respiratory tract.

The Human Connectome Project and beyond: Initial applications of 300 mT/m gradients

The engineering of a 3T human MRI scanner equipped with 300mT/m gradients – the strongest gradients ever built for an in vivo human MRI scanner – was a major component of the NIH Blueprint Human Connectome Project (HCP). This effort was motivated by the HCP's goal of mapping, as completely as possible, the macroscopic structural connections of the in vivo healthy, adult human brain using diffusion tractography. Yet, the 300mT/m gradient system is well suited to many additional types of diffusion measurements. Here, we present three initial applications of the 300mT/m gradients that fall outside the immediate scope of the HCP. These include: 1) diffusion tractography to study the anatomy of consciousness and the mechanisms of brain recovery following traumatic coma; 2) q-space measurements of axon diameter distributions in the in vivo human brain and 3) postmortem diffusion tractography as an adjunct to standard histopathological analysis. We show that the improved sensitivity and diffusion-resolution provided by the gradients are rapidly enabling human applications of techniques that were previously possible only for in vitro and animal models on small-bore scanners, thereby creating novel opportunities to map the microstructure of the human brain in health and disease.