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According to Ayurveda Visha means the substance which causes Vishada (sorrow, depression). Drugs which act against toxic substances are called as Vishaghna (Antitoxic). Acharya Charaka had described ten drugs having anti-toxic property in Vishaghna Mahakashaya. It includes Haridra (Curcuma longa), Manjishtha (Rubia Cardifolia), Suvaha (Pluchea Lanceolata), Sukshma Ela (Elettaria Cardamomum), Palindee (Operculina Turpethum), Chandan (Santalum Album), Kataka (Strychnos Potatorum), Shireesh (Albizzia lebbeck), Sinduvara (Vitex Nigundo), and Shlemantaka (Cordia Dichotoma). Sukshma Ela is one of them. The main objective of this article is to discuss the pharmacological properties and therapeutic uses of Sukshma Ela on allergic skin reactions i.e. Urticaria. This article highlights on those pharmacological properties of Ela which can be used in various allergic skin reactions i.e. Urticaria.

Extensive surveys were undertaken across Kerala during 2018-22, to explore the diversity of aphid fauna associated with different flora. The study recorded 25 species of aphids under 17 genera in four tribes belonging to four subfamilies of Aphididae. Aphidinae was the predominant subfamily, followed by Greenideinae, Hormaphidinae and Eriosomatinae. Aphidinae was represented by two tribes, Aphidini and Macrosiphini. The tribes Aphidini (Aphis (L.), Rhopalosiphum Koch, Hysteroneura Davis and Melanaphis van der Goot) and Macrosiphini (Pentalonia Coquerel, Brevicoryne van der Goot, Macrosiphoniella del Guercio, Macrosiphum Passerini, Metopolophium Mordvilko, Acyrthosiphon Mordvilko, Uroleucon Mordvilko and Aulacorthum Mordvilko) were represented by four and eight genera, respectively. The subfamily Greenideinae was represented by two genera in a single tribe (Greenideini); Hormaphidinae represented by two genera in a single tribe (Cerataphidini), while only one genus in a single tribe (Eriosomatini) was recorded in the subfamily Eriosomatinae. The genus Aphis was the most diverse with five species. Aphis gossypii Glover exhibited a wider host range with 20 host plants. The study records new distribution of three aphid species from south India and seven species from Kerala.

The demand for hydrogen is likely to grow as it could potentially be a suitable alternative for fossil fuels. Hydrogen is used extensively for a variety of purposes in major industries, such as refineries for hydro- desulphurization and hydrocracking operations. In ammonia production it reacts with nitrogen to produce ammonia, in metallic ore reduction, hydrogen is commercially used to procure tungsten from its ore, and various other applications. Hydrogen being a colorless, odorless, and tasteless gas, which is flammable, can’t be identified or recognized by the human senses, and needs to be detected by other means so that one could quantify its concentration if present. There is a need to estimate and determine the concentration of hydrogen so that it can help prevent any potential hazards. This article gives an insight on all the hydrogen sensors that are available in the market, a comparison between all the different hydrogen sensors with different attributes, such as stability, sensitivity, response time, etc., and the different manufacturers who produce this sensor.

Purposive sampling surveys were conducted in homestead gardens and ornamental nurseries at different localities of Thrissur and Ernakulam districts during 2017-19 to study the diversity of spider mites associated with ornamental plants. Mite samples collected from rose, adenium, gerbera, marigold, chrysanthemum, balsam, cock’s comb, orchid, cassia, cairo morning glory, zinnia, bauhinia, crape jasmine and pinto peanut were maintained as isoline cultures in the laboratory. Male and female specimens from isolines were slide mounted for morphological characterisation and identification. The results revealed eight species belonging to three genera viz., Tetranychus truncatus Ehara, T. okinawanus Ehara, T. urticae Koch, T. fijiensis Hirst, T. neocaledonicus Andre, T. marianae McGregor, Eutetranychus orientalis Klein and Oligonychus biharensis Hirst. The study recorded new hosts for the alien species, T. okinawanus and T. truncatus which suggests the potential of these species to turn invasive in Kerala’s ecosystems. The spider mite fauna on rose was more diverse, indicating the need for imposing strict quarantine regulations. The study observed T. marianae as a new record from Kerala.

In contrast to premalignant lesions, ErbB2-initiated tumors in wildtype and ARF-null mammary glands exhibit comparable senescence and DDR.

Loss of ARF does not impact proliferation, autophagy, or DNA damage complex formation.

Loss of ARF results in diminished p53-function and impaired senescence, but does not impact apoptosis.

p16 plays a minor role in mediating senescence in the mammary gland.

ARF-loss does not affect mammary gland development or rate of infection.

Loss of p53 does not diminish DDR following ErbB2 activation.

Acorus calamus (Acoraceae) also known as sweet flag in Indian traditional medicine is generally used for treatment of various ailments like cough, fever, bronchitis, inflammation, depression, tumours, haemorrhoids, skin diseases, insomnia, hysteria, epilepsy, and loss of memory. Asarone is a chemical compound of the phenylpropanoid class found in plants such as Acorus and Asarum. There are two isomers, α (trans) and β (cis). Alpha-asarone is potentially toxic compared to beta-asarone and hence pharmacological elucidation of beta-asarone is wide. Beta-asarone due to its blood brain barrier crossing property it is well elucidated for potential neuroprotective effect. The beneficial properties of beta-asarone are attributed to molecular pathways of endoplasmic reticulum stress, autophagy and synaptogenesis through IRE1/XBP1 ER stress pathway, mitochondrial ASK1/MKK7/JNK pathway, CaMKII/CREB/Bcl-2 expression and PERK/CHOP/Bcl-2/Beclin-1 pathways. The memory enhancing property of beta-asarone is said to be due to beclin dependent autophagy by PI3K/AKT/mTOR pathway. The aim of this review is to highlight the neuroprotective role of beta asarone in terms of neuroinflammation, apoptosis, neurogenesis and autophagy with special emphasis on two neurodegenerative disorders Parkinson's disease and Alzheimer's disease along with its beneficial property in elucidating synaptic plasticity and neurogenesis. Further research on toxicity and pharmacokinetics of beta-asarone are much needed to bring this potential compound into therapeutic use.

We report a case of a huge congenital pulmonary airway malformation (CPAM) that was referred as congenital diaphragmatic hernia (CDH). Initial ultrasound evaluation revealed a huge cystic lesion with septations, in the thorax, causing mediastinal shift and compression effects, suggesting the possibility of a thoracic lymphangioma, or bowel herniation with obstruction. A fetal magnetic resonance imaging reported possible bowel herniation through a posterior defect in the diaphragm, with volvulus, reinforcing the diagnosis of CDH. It was only on autopsy and subsequent histopathology examination that the diagnosis of a rare variant of CPAM–nonmucinous papillary type, could be made. To the best of our knowledge, a CPAM this huge has not been reported prenatally at this gestation. We recommend considering the potential diagnosis of CPAM in any thoracic cystic irrespective of its size or appearance.

This abstract was withdrawn by the authors. Citation Format: Rinkenbaugh AL, Sinha VC, Zhang X, Shao J, Piwnica-Worms H. Withdrawn [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-07-03.

There is an unmet clinical need for stratification of breast lesions as indolent or aggressive to tailor treatment. Here, single-cell transcriptomics and multiparametric imaging applied to a mouse model of breast cancer reveals that the aggressive tumor niche is characterized by an expanded basal-like population, specialization of tumor subpopulations, and mixed-lineage tumor cells potentially serving as a transition state between luminal and basal phenotypes. Despite vast tumor cell-intrinsic differences, aggressive and indolent tumor cells are functionally indistinguishable once isolated from their local niche, suggesting a role for non-tumor collaborators in determining aggressiveness. Aggressive lesions harbor fewer total but more suppressed-like T cells, and elevated tumor-promoting neutrophils and IL-17 signaling, disruption of which increase tumor latency and reduce the number of aggressive lesions. Our study provides insight into tumor-immune features distinguishing indolent from aggressive lesions, identifies heterogeneous populations comprising these lesions, and supports a role for IL-17 signaling in aggressive progression., The classification of breast lesions as indolent or aggressive to tailor treatment is crucial. Here, the authors use single-cell transcriptomics and multiparametric imaging of a breast cancer mouse model, report distinct tumor-immune features for the two types of lesions, and suggest the role of IL-17 signaling in disease progression.

Escherichia coli is a Gram-negative bacterium that colonises the human intestine and virulent strains can cause severe diarrhoeal and extraintestinal diseases. The protein SslE is secreted by a range of pathogenic and commensal E. coli strains. It can degrade mucins in the intestine, promotes biofilm maturation and it is a major determinant of infection in virulent strains, although how it carries out these functions is not well understood. Here, we examine SslE from the commensal E. coli Waksman and BL21 (DE3) strains and the enterotoxigenic H10407 and enteropathogenic E2348/69 strains. We reveal that SslE has a unique and dynamic structure in solution and in response to acidification within mature biofilms it can form a unique aggregate with amyloid-like properties. Furthermore, we show that both SslE monomers and aggregates bind DNA in vitro and co-localise with extracellular DNA (eDNA) in mature biofilms, and SslE aggregates may also associate with cellulose under certain conditions. Our results suggest that interactions between SslE and eDNA are important for biofilm maturation in many E. coli strains and SslE may also be a factor that drives biofilm formation in other SslE-secreting bacteria.

Shifts in tumor cell phenotype in response to selective pressures (i.e. changing microenvironments, drug treatments) pose one of the biggest obstacles to successful breast cancer therapies. Phenotypically diverse breast tumor and stroma subpopulations, and interactions between them that alter tumor cell biology, represent unique and spatially distinct niches. We hypothesize that localized neighborhoods of breast tumor cells possess specialized phenotypes that mediate chemoresistance and represent novel therapeutic vulnerabilities. In order to assess these potential phenotypes, we utilized imaging mass cytometry (IMC), a highly multiplexed imaging modality that allows simultaneous measurement of 30-40 antigens while retaining the spatial architecture of the cancer tissue. We constructed an IMC antibody panel that combines markers for tissue architecture, tumor and stromal cell phenotyping, and signaling pathway activation. IMC was applied to patient-derived xenograft (PDX) models of triple negative breast cancer (TNBC).Our TNBC PDX collection was established from tumors obtained before and after neoadjuvant Adriamycin and cyclophosphamide (AC). IMC analysis of 18 PDX models representing eight patients revealed that stromal cell phenotypes were generally shared between all models, but tumor cell phenotypes were largely patient-specific. While every model was comprised primarily of a few major tumor cell phenotypes, we noted that each case also harbored several minor, unique populations, suggesting that specialized neighborhoods may exist within the tumor mass. Comparison of paired PDX models showed a wide range of phenotypic responses to chemotherapy, ranging from stable tumor composition to widespread changes in tumor phenotypes. These phenotypic changes arose despite relatively consistent genomic architecture. Vimentinhi fibroblasts were present more often in post-AC models, while SMAhi fibroblasts were unchanged after treatment. Comparison of pre-/post-AC PDX pairs revealed spatially constrained MAPK activation emerged after treatment. To capture acute changes in tumor phenotype, we treated treatment-naïve PDX models with AC and evaluated tumors by IMC. As tumors regressed and then regrew, we identified novel phenotypic shifts, again including increased MAPK signaling localized to discrete neighborhoods, suggesting this property may be a common feature of chemoresistant TNBC. Analysis of adjacent cells revealed seven distinct neighborhoods, and ongoing work is aimed at determining whether these neighborhoods are altered in response to chemotherapy treatment. Taken together, our findings suggest that distinct tumor phenotypes arise following treatment. Our goal is to determine whether these unique phenotypic niches functionally contribute to chemoresistance and if disruption of these niches enhances chemosensitivity. Citation Format: Amanda L. Rinkenbaugh, Vidya C. Sinha, Pankaj Singh, Yuan Qi, Jiansu Shao, Xiaomei Zhang, Gloria V. Echeverria, W. Fraser Symmans, Stacy L. Moulder, Helen Piwnica-Worms. Analysis of spatiotemporal phenotypic heterogeneity in chemoresistant triple negative breast cancer using imaging mass cytometry [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1595.

Ductal carcinoma in situ (DCIS) is a non-invasive proliferative growth in the breast that serves as a non-obligate precursor to invasive ductal carcinoma. The widespread adoption of screening mammography has led to a steep increase in the detection of DCIS, which now comprises approximately 20% of new breast cancer diagnoses in the United States. Interestingly, the intratumoral heterogeneity (ITH) that has been observed in invasive breast cancers may have been established early in tumorigenesis, given the vast and varied ITH that has been detected in DCIS. This review will discuss the intratumoral heterogeneity of DCIS, focusing on the phenotypic and genomic heterogeneity of tumor cells, as well as the compositional heterogeneity of the tumor microenvironment. In addition, we will assess the spatial heterogeneity that is now being appreciated in these lesions, and summarize new approaches to evaluate heterogeneity of tumor and stromal cells in the context of their spatial organization. Importantly, we will discuss how a growing understanding of ITH has led to a more holistic appreciation of the complex biology of DCIS, specifically its evolution and natural history. Finally, we will consider ways in which our knowledge of DCIS ITH might be translated in the future to guide clinical care for DCIS patients.

Introduction The greater wing of sphenoid presents various foramina, of which the foramen ovale is one important foramen through which advanced surgical therapeutic and diagnostic procedures related to the middle cranial fossa are performed. Materials and Methods A total of 40 dried adult skulls of unknown gender and age, obtained from the Department of Anatomy of the JSS medical College, Mysuru, Kamakata, India. The length and the width of the foramen ovale were measured using digital sliding calipers (tiny deal 150 mm SS digital caliper with LCD display, Kristeel-Shimwa industries, Bombay, India). Results The mean length of the foramen ovale was 0.745 ± 0.31 cm on the right side (RS), and 0.68 ± 0.15 cm on the left side (LS). The mean width was 0.6 ± 0.17 cm on the RS, and 0.56 ± 0.14 cm on the LS. Conclusion The knowledge of variations in the length and breadth of the foramen ovale is of immense importance in neurosurgery during various invasive surgical procedures, such as percutaneous trigeminal rhizotomy, and in the biopsy of cavernous sinus tumors and of Meckel cave lesions.

Objective To describe the rapid conceptualization and implementation of an interdisciplinary epilepsy clinic for children with drug-resistant epilepsy (DRE) at Arkansas Children's Hospital (ACH) during the COVID 19 pandemic. Methods Focusing on care design and care coordination for children with DRE, multiple stakeholder groups decided to implement a clinic after the systematic rating of constructs present in a theoretical meta-analytic framework. Based on the projected success, the new interdisciplinary clinic (composed of an epileptologist, a neurosurgeon, and a neuropsychologist and coordinated by a full-time nurse) was established. Clinic operations were further refined through discussions with patients, families, and care providers. We collected data retrospectively (August 2020 to June 2021) to determine referral patterns, clinic scheduling metrics, patient characteristics, clinical recommendations, and epilepsy quality metrics. Results Of the 32 Consolidated Framework for Implementation Research constructs assessed, 24 were positively rated to predict a high probability of successful implementation of the clinic. For approximately 100 patient visits, appearance and usage rates were >75%, yielding a clinic utilization rate of approximately 60%. Among 76 unique patients (average age of 12 years, 60% focal epilepsy), 39 patients (51.3%) were deemed eligible for epilepsy surgery evaluation. The majority of the patients (53.9%) were advised for additional diagnostic testing, and 31.6% of patients were scheduled for vagus nerve stimulation. More patients (33%) had changes in their existing anti-seizure medication (ASM) regimen rather than an addition of a new ASM (7.9%). Standardized epilepsy quality measures showed >80% to 90% adherence in 3 (reproductive counseling, depression and anxiety screening, documentation of seizure frequency) out of 4 metrics. Significance This is the first study to show that an interdisciplinary clinic can be a valuable attribute of care models in high-need children with DRE by enabling comprehensive one-stop service for diagnostic evaluation, surgical consideration, and brief assessment of psychiatric comorbidities without compromising consensus-based best practices.

Infrastructural development is crucial for regional trade integration. This study investigates the extent of regional trade interdependence of the Association of Southeast Asian Nations (ASEAN) and its nexus with infrastructure. We selected 13 countries-ASEAN and three other countries (India, the People's Republic of China, and Japan) for the period 1990 to 2018. Using trade network analysis and the panel fixed-effect method, our findings show that: (1) the regional integration in the region is strong and the countries are well interconnected; (2) there is no "center-periphery" structural difference in the region; (3) there are, however, emerging trade-intensive paths among a few countries in the region, which implies the emergence of "exports hubs" and "suppliers of intermediate goods"; (4) the nexus between the infrastructure index and the trade connectivity are strong; (5) shipping and air transport developments have improved the trade connectivity; (6) trade costs (barriers) affect the trade connectivity negatively; and (7) foreign direct investment and trade agreements have helped in improving ASEAN's trade connectivity.

The relationship between ATR/Chk1 activity and replication stress, coupled with the development of potent and tolerable inhibitors of this pathway, has led to the clinical exploration of ATR and Chk1 inhibitors (ATRi/Chk1i) as anticancer therapies for single-agent or combinatorial application. The clinical efficacy of these therapies relies on the ability to ascertain which patient populations are most likely to benefit, so there is intense interest in identifying predictive biomarkers of response. To comprehensively evaluate the components that modulate cancer cell sensitivity to replication stress induced by Chk1i, we performed a synthetic-lethal drop-out screen in a cell line derived from a patient with triple-negative breast cancer (TNBC), using a pooled barcoded shRNA library targeting ~350 genes involved in DNA replication, DNA damage repair, and cycle progression. In addition, we sought to compare the relative requirement of these genes when DNA fidelity is challenged by clinically relevant anticancer breast cancer drugs, including cisplatin and PARP1/2 inhibitors, that have different mechanisms of action. This global comparison is critical for understanding not only which agents should be used together for combinatorial therapies in breast cancer patients, but also the genetic context in which these therapies will be most effective, and when a single-agent therapy will be sufficient to provide maximum therapeutic benefit to the patient. We identified unique potentiators of response to ATRi/Chk1i and describe a new role for components of the cytosolic iron–sulfur assembly (CIA) pathway, MMS19 and CIA2B-FAM96B, in replication stress tolerance of TNBC.

Aims. Epithelial mesenchymal transition (EMT) is a crucial process for acquisition of malignant phenotype, aggressiveness, and metastatic capacity in neoplasms. It is characterized by loss of epithelial markers and gain of mesenchymal markers. Studies on EMT and its potential association with the histological grading are sparse in oral squamous cell carcinoma (OSCC). This study aims to evaluate the expression of EMT-associated proteins—E-cadherin, β-catenin, and N-cadherin—in different grades of OSCC. Methodology. In all, 60 cases of OSCC further subdivided into 20 cases each of well-, moderately, and poorly differentiated OSCCs were stained immunohistochemically with E-cadherin, β-catenin, and N-cadherin antibodies. The differences in the expression were evaluated using χ2 and Fisher exact tests, whereas Spearman’s correlation was used to analyze the correlation between the markers. Results. A reduced E-cadherin expression noted in 40% of the OSCCs was associated with reduced β-catenin expression in 66.6% of the cases and increase in the expression of mesenchymal N-cadherin seen in 80% of cases. This expression pattern demonstrated a significant association with histological grades. A membrane to cytoplasmic shift of E-cadherin (73.3%) and β-catenin (78.3%) increased with histological grade. A negative correlation was observed with the E-cadherin and N-cadherin localization, though it did not reach statistical significance. Conclusion. OSCC tissues had high levels of EMT phenotype as compared with the normal oral mucosa. This phenotype was characterized by reduced E-cadherin and β-catenin expression and overexpression of N-cadherin. Aberrant localization of the studied proteins was a hallmark for depicting EMT.

Shifts in tumor phenotype in response to selective pressures (i.e. changing microenvironments, drug treatments) pose one of the biggest obstacles to successful cancer therapies. Phenotypically diverse tumor and stroma subpopulations, and interactions between them that alter tumor biology, represent unique signaling niches. We hypothesize that these localized neighborhoods of breast tumor cells possess specialized phenotypes that mediate chemoresistance and represent novel therapeutic vulnerabilities. In order to assess these potential phenotypes, we utilized imaging mass cytometry (IMC), a highly multiplexed imaging modality that allows simultaneous measurement of 30-40 antigens while retaining the spatial architecture of the cancer tissue. We have constructed an IMC antibody panel that combines markers for tissue architecture, tumor and stromal cell phenotyping, and signaling pathway activation. IMC was applied to a collection of patient-derived xenograft (PDX) models of triple negative breast cancer (TNBC). Our PDX collection was established from patient tumors obtained before and after neoadjuvant chemotherapy. IMC analysis of 18 PDX models representing eight patients revealed that stromal cell phenotypes were generally shared between all models, but tumor cell phenotypes were largely patient-specific. While every model was comprised primarily of a few major tumor cell phenotypes, we noted that each case also harbored several minor, unique populations, suggesting that specialized neighborhoods may exist within the tumor mass. Sequential PDX models showed a wide range of phenotypic responses after chemotherapy, ranging from stable tumor composition to widespread changes in tumor phenotypes. Importantly, comparison of multiple pre-/post-chemotherapy PDX pairs revealed spatially constrained MAPK activation emerging after treatment. To capture acute changes in tumor phenotype, we treated animals bearing PDX tumors with chemotherapy and evaluated tumors by IMC. As tumors regressed and then regrew, we identified novel phenotypic shifts, again including increased MAPK signaling localized to discrete neighborhoods, suggesting this phenomenon may be a common feature of chemoresistant TNBC. Taken together, our findings suggest that distinct tumor phenotypes arise following treatment. Ongoing work is examining the spatial arrangement of these cell types to determine their local niche compositions, and then correlating these findings with clinical features. Our goal is to determine whether these unique signaling niches functionally contribute to chemoresistance and if disruption of these niches enhances chemosensitivity. Citation Format: Amanda L. Rinkenbaugh, Vidya C. Sinha, Jiansu Shao, Xiaomei Zhang, Gloria V. Echeverria, W. Fraser Symmans, Stacy L. Moulder, Helen Piwnica-Worms. Analysis of spatiotemporal phenotypic heterogeneity in chemoresistant triple negative breast cancer using imaging mass cytometry [abstract]. In: Proceedings of the AACR Virtual Special Conference on Tumor Heterogeneity: From Single Cells to Clinical Impact; 2020 Sep 17-18. Philadelphia (PA): AACR; Cancer Res 2020;80(21 Suppl):Abstract nr PR01.

Introduction: Ductal carcinoma in situ is a non-invasive lesion of the breast that comprises ~20% of newly diagnosed breast cancers in the United States. Prospectively distinguishing indolent from aggressive lesions has been a major clinical challenge. To begin addressing these challenges, we use an experimentally tractable mouse model of breast cancer in which invasive (but rarely in situ) lesions display architectural and morphological abnormalities while also being heavily immune-infiltrated. Methods: To explore the tumor-stroma niches that evolve as breast cancer progresses, we are undertaking single cell RNA sequencing and imaging mass cytometry to identify tumor and immune cells present within in situ (early) versus invasive (advanced) lesions, and to characterize shifts in phenotype and tumor-stroma niches that may occur as breast cancer transitions from in situ to invasive disease. Results: Single cell analyses revealed major cell populations within the tumor compartment of both early and advanced disease, including luminal- and basal-like populations, as well as an additional minor population that appears to represent a unique intermediate between luminal and basal states. Early stage lesions were comprised primarily of the major luminal population, whereas advanced lesions exhibited a significant expansion of intermediate and basal populations, suggesting that breast cancer cells may undergo a transition from luminal-like to basal-like phenotype during progression to invasive disease. Analysis of the immune compartments of early versus advanced disease also revealed that the relative neutrophil/MDSC/S100A8+ population increased as lesions advanced, while B cells, NK cells, and some T cells decreased. Furthermore, the immune populations of advanced lesions exhibited widespread upregulation of IL-17 targets, suggesting that the IL-17 pathway may drive shifts in the immune microenvironmental to support disease progression, particularly the increase in neutrophils/MDSCs. Additionally, given a previously reported role of IL-17 to alter epithelial stemness, increased IL-17 signaling in advanced lesions may also be driving the potential luminal-to-basal transition of malignant cells. Conclusion: Taken together, these findings suggest that a subset of malignant epithelial cells may transition from luminal-like to basal-like cells, and that this transition may be driven and/or maintained by a commensurate increase in IL-17 signaling that both directly impacts tumor cells and also establishes a tumor-supportive microenvironment. Citation Format: Vidya C. Sinha, Mingchu Xu, Amanda L. Rinkenbaugh, Xinhui Zhou, Xiaomei Zhang, Helen Piwnica-Worms. Single-cell evaluation to identify tumor-stroma niches driving the transition from in situ to invasive breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2822.

Introduction: Imaging mass cytometry (IMC) enables highly multiplexed assessment of more than 30 proteins at a time for the quantification of cell phenotypes within the tumor microenvironment of formaldehyde fixed-paraffin embedded (FFPE) tumor tissue. This methodology provides a deep understanding of the immune-contexture while preserving their spatial information in regards to malignant cells. We report a detailed workflow for optimization of a 27-plex immune-oncology (IO) panel, immune-phenotyping, and spatial analysis of tumor tissue. Methods: IO-panel includes 27 antibodies targeting lymphoid (CD3e, CD4, CD8a, CD19, CD45RO, LAG3, ICOS, Granzyme-B), myeloid (CD11b, CD14, CD33, CD68, CSF1R, IDO-1), immune-regulatory (HLA-DR, OX-40, VISTA, TIM-3, CD73, B7-H3), epithelial (Cytokeratin), proliferative (Ki-67), and constitutive (GAPDH, NaKATPase, Vimentin, aSMA, Histone H3) markers that were selected by singlet automated chromogenic immunohistochemistry (IHC) staining. BSA-free formulation from the same clones of antibodies were isotope-metal conjugated and manual indirect immunofluorescence (IF) staining was used to confirm the stability of metal-tagged antibodies. The optimization by singlet-IF and multiplexed-IMC staining was performed with normal and malignant tissues (4mm core TMA with tonsil, placenta, prostate, breast carcinoma, ovarian carcinoma, and endometrioid carcinoma tissues). All images from IHC (HALO software), indirect IF (InForm software), and IMC (MCD viewer software) slides were evaluated by two pathologists. Cell densities and spatial analysis of a breast carcinoma core was performed using HALO software. Results: All antibodies included in the panel were individually optimized by IHC under the same protocol conditions. A significant correlation of optimal antibody concentrations where observed across the different staining techniques (IHC vs IF, r=0.51; IHC vs IMC, r= 0.54; IF vs IMC, r=0.992; all p values Conclusions: IMC is a powerful platform for highly multiplexed detection of protein and cell phenotypes quantification in FFPE samples. Our developed IMC panel and digital image analysis of tissues allow the quantification of immune cells and spatial analysis of the tumor tissue microenvironment. Citation Format: Pedro Rocha, Luisa Solis, Edwin Parra, Ou Shi, Barbara Mino, Vidya C. Sinha, Amanda Rinkenbaugh, Auriole Tamegnon, Mak Duncan, Wei Lu, Mei Jiang, Lakshimi Kakarala, Jared K. Burks, Helen Piwnica-Worms, Cara Haymacker, Michael Teztlaff, Ignacio Wistuba, Alejandro Francisco-Cruz. Immuno-oncology panel optimization for imaging mass cytometry and digital image analysis of tumor tissues [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2679.

Tumors are increasingly appreciated as complex ecosystems, wherein functional interactions between tumor subclones, as well as components of the microenvironment, contribute to progression and drug resistance. While some studies have focused on soluble factors that mediate these interactions, little is known about the communication between physically neighboring tumor subclones (and their microenvironment). To investigate the nature and impact of such localized interactions, we are assessing the activation status of key cancer signaling pathways in neighboring tumor cells; importantly, we are characterizing these features at the single cell level within physically intact tumors. Triple negative breast cancer (TNBC) exhibits a high degree of intratumor heterogeneity, which has contributed to a lack of effective targeted therapy options. Chemotherapy remains the standard of care; however approximately half of patients have substantial residual disease following chemotherapy, which is associated with a high risk of recurrence. Our objective is to spatially define signaling heterogeneity using patient-derived xenograft (PDX) models before and after chemotherapy treatment, to determine if spatially-defined signaling niches drive chemoresistance in TNBC. We hypothesize that neighborhoods of cells possess specialized phenotypes that mediate chemoresistance, and when disrupted, will inhibit the growth of chemoresistant tumors. We are employing imaging mass cytometry (IMC),a next-generation immunostaining approach that allows for simultaneous measurement of 30-40 biomarkers while retaining the spatial organization of the sample.Wehave constructed an IMC panel of antibodies that combines markers for tissue architecture, tumor and immune cell phenotyping, and signaling pathway activation. Our PDX collection features sequential pairs derived from biopsies taken before and after chemotherapy treatment. IMC of the pre-/post-chemotherapy pairs revealed spatial patterns of pathway activation that emerged following treatment, including increases in PI3K/mTOR and localized MAPK signaling. To complement these studies, we treated PDX models with chemotherapy and analyzed tumors via IMC throughout the course of treatment. Again, we observed the emergence of increased MAPK signaling, localized to discrete neighborhoods within the tumor. Taken together, our findings suggest that unique signaling niches arise following treatment. Our goal is to now determine whether these signaling niches functionally contribute to chemoresistance and if disruption of these niches can inhibit the growth of chemoresistant disease. Citation Format: Amanda L. Rinkenbaugh, Vidya C. Sinha, Gloria V. Echeverria, Xiaomei Zhang, Jiansu Shao, W. Fraser Symmans, Stacy L. Moulder, Helen Piwnica-Worms. Analysis of spatiotemporal phenotypic heterogeneity in chemoresistant triple negative breast cancer using imaging mass cytometry [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1513.

Now a days notice board has become an important thing in institutes/organization or public places like railway stations, bus stands and hospitals. But to use the paper notices stacked on a notice board is a time taking and expensive process and there is wastage lot of time, paper and labour. The notice board is used to display the information in an effective way to the people, but to update the messages instantly is not easy on the notice board. In this paper we have proposed and implemented an advanced Hi-Tech wireless Notice Board. An Android application of smart phones or tablet which is connected to Bluetooth (HC-0S) can be used to display the latest information by enhancing the system. A low cost programmed microcontroller (arduino uno) is used at the receiver to receive and display messages in the LCD display. Instead of sticking notices manually on a notice board the authorized user can speak and the verbalized voice is sent through a Bluetooth and displayed on the LCD screen. In this project we are using a flexible and most successful speech recognize technique called as Hidden Markov Model(HMM) to recognize the speech for voice and then that recognized sampled speech voice are processed identified as a verbalized text done by connecting to a google's server via internet. And the speech is inputted from a microphone and only an English message has taken as an input for our application and it is online speech to text conversion.

Breast cancers exhibit high intertumoral heterogeneity in genetic alterations as well as histopathological and other phenotypic characteristics. The contribution of the initiating oncogenic mutation to tumor phenotype remains controversial, largely due to the technical difficulties in delivering genetic alterations into well-defined subsets of mammary epithelial cells. To examine how different initiating oncogenes drive tumor phenotype, we somatically delivered two oncogenes (ErbB2, PyMT) into a narrow and distinct subset of the mouse mammary epithelium defined by the expression of the progenitor marker keratin 6a (Krt6a), and compared the phenotypes of the resulting mammary tumors. While PyMT-induced tumors were well-differentiated and displayed glandular and papillary features, ErbB2-induced tumors were poorly differentiated and exhibited epithelial-to-mesenchymal transition as well as β-catenin activation. These in vivo data demonstrate that the initiating oncogene plays a key role in driving mammary tumor phenotype.

In response to oncogene activation and oncogene-induced aberrant proliferation, mammalian cells activate apoptosis and senescence, usually via the p53–ARF tumor-suppressor pathway. Apoptosis is a known barrier to cancer and is usually downregulated before full malignancy, but senescence as an anticancer barrier is controversial due to its presence in the tumor environment. In addition, senescence may aid cancer progression via releasing senescence-associated factors that instigate neighboring tumor cells. Here, it is demonstrated that apoptosis unexpectedly remains robust in ErbB2 (ERBB2/HER2)-initiated mammary early lesions arising in adult mice null for either p53 or ARF. These early lesions, however, downregulate senescence significantly. This diminished senescence response is associated with accelerated progression to cancer in ARF-null mice compared with ARF–wild-type mice. Thus, the ARF–p53 pathway is dispensable for the apoptosis anticancer barrier in the initiation of ErbB2 breast cancer, the apoptosis barrier alone cannot halt mammary tumorigenesis, and senescence is a key barrier against carcinogenesis. Implications: Findings in this relevant mouse model of HER2-driven breast cancer suggest that effective prevention relies upon preserving both ARF/p53–independent apoptosis and ARF/p53–dependent senescence. Mol Cancer Res; 13(2); 231–8. ©2014 AACR.

Lung Cancer is a disease can be defined as uncontrolled cell growth in tissues of the lung. If we detect the Lung Cancer in its early stage, then that could be the key of its cure. In this work the non-invasive methods are studied for assisting in nodule detection. It supplies a Computer Aided Diagnosis System (CAD) for early detection of lung cancer nodules from the Computer Tomography (CT) images. CAD system is the one which helps to improve the diagnostic performance of radiologists in their image interpretations. The main aim of this technique is to develop a CAD system for finding the lung cancer using the lung CT images and classify the nodule as Benign or Malignant. For classifying cancer cells, SVM classifier is used. Here, image processing techniques have been used to de-noise, to enhance, for segmentation and edge detection of an image is used to extract the area, perimeter and shape of nodule. The core factors of this research are Image quality and accuracy.

Background: Liver being the largest organ in the body receives blood supply from hepatic artery (25%) and portal vein (75%) and drained by three major hepatic veins opening into inferior vena cava. The dimensions of hepatic veins are quite essential during portosystemic shunting procedure and radiological interventions. The presence of variants of major hepatic veins is of prime importance to evaluate donor and recipient in living donor liver transplantation. Objectives: The objectives of the study were (1) to evaluate the ramification pattern and morphometry of major hepatic veins of liver by cast method and (2) to study the clinical implications of the hepatic vein variations. Materials and Methods: Forty liver specimens (22 males and 18 females) aged between 30 and 60 years were obtained from the Department of Forensic Medicine of JSS Medical College and Mysore Medical College. The silicone gel was injected into vena cava and pushed thoroughly into the ostia of hepatic veins. The drainage patterns of hepatic veins were noted and measurement of hepatic veins was done using thread and measuring scale. Results: The present study ramification pattern of middle and left hepatic veins showed Type 1 pattern in 24 specimens (58%), Type 2 in 12 specimens (40%), and Type 3 in 5 specimens (12%). There was a significant difference with P < 0.05 in all the parameters among males and females except the length and diameter of common trunk. Conclusion: Thorough knowledge of hepatic vein variants and its morphometry is essential during imaging procedures, anatomic resection of hepatic veins in hepatocellular carcinoma, liver transplantation, and other liver surgeries.

Binding and hydrolysis of ATP is universally required by AAA+ proteins to underpin their mechano-chemical work. Here we explore the roles of the ATPase site in an AAA+ transcriptional activator protein, the phage shock protein F (PspF), by specifically altering the Walker B motif sequence required in catalyzing ATP hydrolysis. One such mutant, the E108Q variant, is defective in ATP hydrolysis but fully remodels target transcription complexes, the RNAP-σ(54) holoenzyme, in an ATP dependent manner. Structural analysis of the E108Q variant reveals that unlike wild-type protein, which has distinct conformations for E108 residue in the ATP and ADP bound forms, E108Q adapts the same conformation irrespective of nucleotide bound. Our data show that the remodeling activities of E108Q are strongly favored on pre-melted DNA and engagement with RNAP-σ(54) using ATP binding can be sufficient to convert the inactive holoenzyme to an active form, while hydrolysis per se is required for nucleic acid remodeling that leads to transcription bubble formation. Furthermore, using linked dimer constructs, we show that RNAP-σ(54) engagement by adjacent subunits within a hexamer are required for this protein remodeling activity while DNA remodeling activity can tolerate defective ATP hydrolysis of alternating subunits.

Tumors are highly heterogeneous populations of cells, and measures of intratumoral heterogeneity (ITH) and diversity correlate with worse prognosis in many cancers, including breast cancer. Emerging studies are highlighting functional interactions between subclones, as well as among subclones and components of the tumor microenvironment. However, these studies have largely focused on soluble factors without interrogating the spatial distribution of subclones defined by activated signaling pathways. Previous work in this area has been severely limited by technical restrictions - existing techniques allowing measurement of many biomarkers simultaneously lose all information about the tissue architecture, while those that do retain spatial information can only assay a handful of markers at once. We will circumvent these limitations by undertaking imaging mass cytometry (IMC), which allows for simultaneous measurement of 30-40 antigens while retaining the spatial organization of the sample. Our objective is to dissect the signaling heterogeneity in tumors from patients and patient-derived xenograft (PDX) models of triple negative breast cancer (TNBC), through two main approaches: (1) characterization of signaling heterogeneity in tissue microarrays of human tumors and PDX models of TNBC with IMC and (2) modeling cell signaling heterogeneity in cell line-based models to determine mechanisms of cell-cell interaction and communication. We have constructed an IMC panel of antibodies that combines markers for tissue architecture, tumor and immune cell phenotyping, and signaling pathway activation. Profiling of a diverse panel of TNBC PDX models captures the heterogeneity of human TNBC. Analysis of distinct regions within individual PDX tumors demonstrate unique compositions of cell phenotypes between the edge and core of the tumor. Our PDX collection also includes sequential pairs derived from biopsies taken before and after chemotherapy treatment. Comparison of the pre-/post-chemotherapy pairs indicates emerging patterns of pathway activation. Interestingly, while the tumor cells from these models exhibit distinct phenotypes, the stromal cells are largely indistinguishable from one another, suggesting that these models are capturing tumor cell-intrinsic changes associated with chemotherapy response. Ultimately, we plan to use these results to identify novel vulnerabilities in subclonal interactions that can be targeted therapeutically in TNBC. Citation Format: Amanda L. Rinkenbaugh, Vidya C. Sinha, Xiaomei Zhang, Jiansu Shao, Helen Piwnica-Worms. Investigating triple negative breast cancer phenotypic heterogeneity of human and patient-derived xenograft samples using imaging mass cytometry [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4708.

Transcription initiation is highly regulated in bacterial cells, allowing adaptive gene regulation in response to environment cues. One class of promoter specificity factor called sigma54 enables such adaptive gene expression through its ability to lock the RNA polymerase down into a state unable to melt out promoter DNA for transcription initiation. Promoter DNA opening then occurs through the action of specialized transcription control proteins called bacterial enhancer-binding proteins (bEBPs) that remodel the sigma54 factor within the closed promoter complexes. The remodelling of sigma54 occurs through an ATP-binding and hydrolysis reaction carried out by the bEBPs. The regulation of bEBP self-assembly into typically homomeric hexamers allows regulated gene expression since the self-assembly is required for bEBP ATPase activity and its direct engagement with the sigma54 factor during the remodelling reaction. Crystallographic studies have now established that in the closed promoter complex, the sigma54 factor occupies the bacterial RNA polymerase in ways that will physically impede promoter DNA opening and the loading of melted out promoter DNA into the DNA-binding clefts of the RNA polymerase. Large-scale structural re-organizations of sigma54 require contact of the bEBP with an amino-terminal glutamine and leucine-rich sequence of sigma54, and lead to domain movements within the core RNA polymerase necessary for making open promoter complexes and synthesizing the nascent RNA transcript.

Our overall goal is to distinguish aggressive from indolent breast cancers by characterizing determinants of breast cancer progression. The detection of early-stage (in situ) neoplastic lesions in the breast has risen dramatically due to widespread mammography. Early detection represents a significant opportunity to intervene and halt the progression of these early lesions to invasive breast cancer. However, our understanding of the natural history of in situ lesions remains limited. Here, we describe our efforts to characterize the microenvironment of DCIS in patient samples and mouse models using a combination of traditional and novel multiplexed immunostaining methods. In particular, we present our early work on applying multispectral fluorescence imaging as well as highly parametric imaging mass cytometry to describe the immune cells surrounding and infiltrating DCIS, with focus on the interaction of these immune cells with each other and the incipient invasive tumor cells. Future studies will be directed at determining if the immune microenvironment of DCIS can identify patients at risk for progression or recurrence so that these patients can be appropriately managed upfront. Citation Format: Vidya C. Sinha, Fei Yang, Amanda L. Rinkenbaugh, Xinhui Zhou, Xiaomei Zhang, Ignacio I. Wistuba, Helen Piwnica-Worms. Multiparametric imaging to evaluate human and mouse breast carcinoma in situ [abstract]. In: Proceedings of the AACR Special Conference: Advances in Breast Cancer Research; 2017 Oct 7-10; Hollywood, CA. Philadelphia (PA): AACR; Mol Cancer Res 2018;16(8_Suppl):Abstract nr B59.

Tumors are highly heterogeneous populations of cells, and measures of intratumoral heterogeneity (ITH) and diversity correlate with worse prognosis in many cancers, including breast cancer. Emerging studies are highlighting functional interactions between subclones, as well as among subclones and components of the tumor microenvironment. However, these studies have largely focused on soluble factors without interrogating the spatial distribution of subclones defined by activated signaling pathways. Previous work in this area has been severely limited by technical restrictions – existing techniques allowing measurement of many biomarkers simultaneously lose all information about the tissue architecture, while those that do retain spatial information can only assay a handful of markers at once. We will circumvent these limitations by undertaking imaging mass cytometry (IMC), which allows for simultaneous measurement of 30-40 antigens while retaining the spatial organization of the sample. Our objective is to dissect the signaling heterogeneity in tumors from patients and patient-derived xenograft (PDX) models of triple negative breast cancer (TNBC), through two main approaches: (1) characterization of signaling heterogeneity in human tumors and PDX models of TNBC through the use of imaging mass cytometry (IMC) and (2) modeling cell signaling heterogeneity in cell line-based models to determine mechanisms of cell-cell interaction and communication. We have constructed an IMC panel of antibodies that combines markers for tissue architecture, cell phenotyping, and signaling pathway activation. Profiling of a panel of TNBC PDX models, including matched sets of pre- and post-chemotherapy treatment, indicates emerging patterns of pathway activation. We will utilize these results to inform functional studies in our isogenic cell line models. As it is becoming more clear that tumors are composed of heterogeneous subclones, it is crucial that we understand the communication between subclones and with components of the microenvironment. Ultimately, we plan to use these results to identify novel vulnerabilities in subclonal interactions that can be targeted therapeutically in TNBC. Citation Format: Amanda L. Rinkenbaugh, Vidya C. Sinha, Xiaomei Zhang, Helen Piwnica-Worms. Functionalizing intratumoral signaling heterogeneity in triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2181.

Summary Inhibition of kinesin-5, a mitotic motor protein also expressed in neurons [1], causes axons to grow faster as a result of alterations in the forces on microtubules (MTs) in the axonal shaft [2–4]. Here, we investigate whether kinesin-5 plays a role in growth-cone guidance. Growth-cone turning requires that MTs in the central (C-) domain enter the peripheral (P-) domain in the direction of the turn. We found that inhibition of kinesin-5 in cultured neurons prevents MTs from polarizing within growth cones and causes them to grow past cues that would normally cause them to turn. We found that kinesin-5 is enriched in the transition (T-) zone of the growth cone and that kinesin-5 is preferentially phosphorylated on the side opposite the invasion of MTs. Moreover, when a growth cone encounters a turning cue, phospho-kinesin-5 polarizes even before the growth cone turns. Additional studies indicate that kinesin-5 works in part by antagonizing cytoplasmic dynein and that these motor-driven forces function together with the dynamic properties of the MTs to determine whether MTs can enter the P-domain. We propose that kinesin-5 permits MTs to selectively invade one side of the growth cone by opposing their entry into the other side.

The phosphatidylinositol 3-kinase-related protein kinases are key regulators controlling a wide range of cellular events. The yeast Tel1 and Mec1·Ddc2 complex (ATM and ATR-ATRIP in humans) play pivotal roles in DNA replication, DNA damage signaling, and repair. Here, we present the first structural insight for dimers of Mec1·Ddc2 and Tel1 using single-particle electron microscopy. Both kinases reveal a head to head dimer with one major dimeric interface through the N-terminal HEAT (named after Huntingtin, elongation factor 3, protein phosphatase 2A, and yeast kinase TOR1) repeat. Their dimeric interface is significantly distinct from the interface of mTOR complex 1 dimer, which oligomerizes through two spatially separate interfaces. We also observe different structural organizations of kinase domains of Mec1 and Tel1. The kinase domains in the Mec1·Ddc2 dimer are located in close proximity to each other. However, in the Tel1 dimer they are fully separated, providing potential access of substrates to this kinase, even in its dimeric form.

Keeping gene transcription in check Transcription of all genes is carried out by RNA polymerase (RNAP). The enzyme is thus a pivotal regulation point for many cell and developmental processes. In bacteria, sigma factors play a vital role in transcription regulation, with σ54 being critical for transcription of many stress response genes. Yang et al. determined the x-ray crystal structure of RNAP bound to σ54, as well as promoter DNA. In the initial inhibited state of the RNAP-σ54 complex, the σ54 blocks the template DNA from entering the RNAP active site and the downstream DNA channel. Science , this issue p. 882

Glucose transporters, such as GLUT-1, mediate the important mechanisms involved in cellular glucose influx, allowing cells to proliferate and survive. The significance of GLUT-1 expression in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) has been less explored, and no study has investigated it in relation to verrucous carcinoma (VC). We evaluated 30 cases each of OED, OSCC, and VC, graded further on the basis of their differentiation, immunohistochemically for GLUT-1 expression, along with 10 specimens of normal oral mucosa (NOM) as controls. In OSCC, GLUT-1 expression increased with the degree of dysplasia and increasing grade (P < 0.001). The expression in VC was predominantly membranous and intense, resembling well differentiated OSCC. This increase of GLUT-1 expression in OSCC along with the degree of dysplasia and the histologic grade reflects the expanding glycolytic response to hypoxia. This is the first study to have revealed prominent GLUT-1 expression in VC, highlighting its inherent metabolic capacity.

Type IV secretion systems (T4SSs) are large multisubunit translocons, found in both gram-negative and gram-positive bacteria and in some archaea. These systems transport a diverse array of substrates from DNA and protein–DNA complexes to proteins, and play fundamental roles in both bacterial pathogenesis and bacterial adaptation to the cellular milieu in which bacteria live. This review describes the various biochemical and structural advances made toward understanding the biogenesis, architecture, and function of T4SSs.

Background: The internal carotid artery supplies the anterior part of the brain, the eye and its appendages and sends branches to the forehead and nose.Carotid canal is a curved channel in the petrous temporal bone through which internal carotid artery enters the skull. Aim: In the present study 20 skulls of unknown sex were considered. The morphometric study of carotid canal was studied by direct bone method and by silicone cast method. The present study aimed to show the differences of measurements by two methods and the silicon cast method is of its kind to measure the length of the carotid canal. Materials and Methods: Carotid canal were measured at three different levels by both methods. Length of carotid canal to bend, diameter at the bend, length from the bend to foramen lacerum. The data was analysed for all the measurements of two methods and comparison of mean and SD at three different levels was done.Significant difference between two methods at three different levels was observed. Independent sample t-test was applied for total length of carotid canal. Results: By observation there is a mean significant difference between two methods in the measurements of carotid canal from lower opening till the angulation on left side of the skull.The study showed that there is a bilateral significant difference in measurements of diameter between two methods. There was no significant difference observed in total length of the canal in both left and right sides. Conclusion: Morphometrical details of the canal are of great significance to neurosurgeons and otologists. Silicon cast method was cost effective and much easier method to study the length and measurements of carotid canal as it exactly depicts the curvature of carotid canal.

Recent studies on cultured neurons have demonstrated that microtubules are transported down the axon in the form of short polymers. The transport of these microtubules is bidirectional, intermittent, asynchronous, and occurs at the fast rate of known motors. The majority of the microtubule mass in the axon exists in the form of longer immobile microtubules. We have proposed a model called 'cut and run', in which the longer microtubules are mobilized by enzymes that sever them into shorter mobile polymers. In this view, the molecular motors that transport microtubules are not selective for short microtubules but rather impinge upon microtubules irrespective of their length. In the case of the longer microtubules, these motor-driven forces do not transport the microtubules in a rapid and concerted fashion but presumably affect them nonetheless. Here, we discuss the mechanisms by which the short microtubules are transported and suggest possibilities for how analogous mechanisms may align and organize the longer microtubules and functionally integrate them with each other and with the actin cytoskeleton.

Portal vein provides about three-fourths of liver's blood supply. Portal vein is formed behind the neck of pancreas, at the level of the second lumbar vertebra and formed from the convergence of superior mesenteric and splenic veins. The purpose of this study is to review the normal distribution and variation, morphometry of portal vein and its branches for their implication in liver surgery and preoperative portal vein embolization. It is also helpful for radiologists while performing radiological procedures. A total of fresh 40 livers with intact splenic and superior mesenteric vein were collected from the mortuary of Forensic Department, JSS Medical College and Mysuru Medical College. The silicone gel was injected into the portal vein and different segments were identified and portal vein variants were noted. The morphometry of portal vein was measured by using digital sliding calipers. The different types of portal vein segmental variants were observed. The present study showed predominant type I in 90% cases, type II 7.5% cases, and type III 2.5% cases. Mean and standard deviation (SD) of length of right portal vein among males and females were 2.096±0.602 cm and 1.706±0.297 cm, respectively. Mean and SD of length of left portal vein among males and females were 3.450±0.661 cm and 3.075±0.632 cm, respectively. The difference in the Mean among the males and females with respect to length of right portal vein and left portal vein was found to be statistically significant (P=0.010). Prior knowledge of variations regarding the formation, termination and tributaries of portal vein are very helpful and important for surgeons to perform liver surgeries like liver transplantation, segmentectomy and for Interventional Radiologists.

In order for growth cones to turn, microtubules from the central domain must preferentially invade the peripheral domain in the direction of the turn. Recent studies suggest that kinesin-5 (also called Eg5 or kif11) suppresses the invasion of microtubules into the peripheral domain on the side of the growth cone opposite the direction of turning. In theory, kinesin-5 could elicit these effects by acting on the microtubules within the peripheral domain itself, by acting on microtubules in the central domain, or in the transition zone between these two domains. In rat neurons expressing kinesin-5, we documented the presence of kinesin-5 in both domains of the growth cone and especially enriched in the transition zone. We then focally inactivated kinesin-5 in various regions of the growth cone, using micro-chromophore-assisted laser inactivation. We found that a greater invasion of microtubules into the peripheral domain occurred when kinesin-5 was inactivated specifically in the transition zone. However, there was no effect on microtubule invasion into the peripheral domain when kinesin-5 was inactivated in the peripheral domain itself or in the central domain. In other experiments, frog growth cones were observed to turn toward a gradient of a drug that inhibits kinesin-5, confirming that asymmetric inactivation of kinesin-5 can cause the growth cone to turn. Finally, expression of a phospho-mutant of kinesin-5 resulted in greater microtubule invasion throughout the peripheral domain and an inhibition of growth cone turning, implicating phosphorylation as a means by which kinesin-5 is regulated in the growth cone.

Childhood germ cell tumors (cGCTs), believed to arise from transformed primordial germ cells by an unknown mechanism, provide a unique model system for investigating cell signaling, pluripotency, and the microenvironment of neoplastic stem cells (NSCs) in vivo. This is the first report of proteomics of cGCTs.Four dysgerminomas (DYSs) and four childhood endodermal sinus tumors (cESTs), resembling self-renewing and differentiating NSCs, respectively, were selected. Proteomic studies were performed by 2-DE, SDS-PAGE, and cLC/MS/MS with protein database searching.2-DE: 9 of 941 spots were differentially regulated with greater than a twofold change in spot volume for at least three of four gels in each group. Two of nine spots had P values for the t-test analysis of comparisons less than 0.001, while the remaining spots had P values from 0.013 to 0.191. Top-ranked proteins were identified in nine of nine spots with 4.0-38% sequence coverage. APOA1, CRK, and PDIA3 were up-regulated in cESTs. TFG, TYMP, VCP, RBBP, FKBP4, and BiP were up-regulated in DYSs. SDS-PAGE: Up-regulation of NF45 and FKBP4 was observed in four of four cESTs and DYSs, respectively. The fold-changes observed correspond with characteristic genetic changes.Differential regulation of FKBP4 and NF45, combined with previous research on immunosuppressant binding, suggests that glucocorticoid receptor signaling merits further investigation in cGCTs and NSCs.

Kinesin-12 (also called Kif15) is a mitotic motor protein that continues to be expressed in developing neurons. Depletion of kinesin-12 causes axons to grow faster, more than doubles the frequency of microtubule transport in both directions in the axon, prevents growth cones from turning properly, and enhances the invasion of microtubules into filopodia. These results are remarkably similar to those obtained in previous studies in which neurons were depleted of kinesin-5 (also called Eg5 or Kif11), another mitotic motor protein that continues to be expressed in developing neurons. However, there are also notable differences in the phenotypes obtained with depleting each of these motors. Depleting kinesin-12 decreases axonal branching and growth cone size, whereas inhibiting kinesin-5 increases these parameters. In addition, depleting kinesin-12 diminishes the appearance of growth-cone-like waves along the length of the axon, an effect not observed with depletion of kinesin-5. Finally, depletion of kinesin-12 abolishes the “waggling” behavior of microtubules that occurs as they assemble along actin bundles within filopodia, whereas inhibition of kinesin-5 does not. Interestingly, and perhaps relevant to these differences in phenotype, in biochemical studies, kinesin-12 coimmunoprecipitates with actin but kinesin-5 does not. Collectively, these findings support a scenario whereby kinesin-12 shares functions with kinesin-5 related to microtubule–microtubule interactions, but kinesin-12 has other functions not shared by kinesin-5 that are related to the ability of kinesin-12 to interact with actin.

Neurons are terminally differentiated cells that do not utilize their microtubules for the formation of mitotic spindles. Instead, microtubules serve as architectural elements within axons and dendrites and also act as railways for the transport of organelles. Molecular motors transport organelles along the microtubule, but can also transport the microtubules themselves and organize them into regionally distinct patterns. Motor and nonmotor microtubule-related proteins can influence the assembly and other properties of the microtubules as well as their interactions with various elements of the cytoplasm. Abnormalities of microtubule-related proteins can give rise to debilitating neuropathies.