Search

Adrenal cystic lymphangiomas are unusual entities with a very low prevalence that are typically discovered incidentally. We describe a 45-year-old male known for hypertension that was referred to our endocrinology department for right adrenal incidentaloma. Seven months prior referral, he presented right hypochondrium pain, vomiting and nausea and imaging studies included a computer tomography that revealed an irregular-shaped right adrenal lesion (42 × 23 mm), with scattered calcifications in the medial branch and heterogenous contrast enhancement and contrast-enhanced computer tomography showed an absolute wash-out of 28% and hyperattenuating areas (>90 HU). Physical examination was unremarkable. Hormonal studies were all within normal range. Due to the findings of imaging studies and lack of biochemical findings a presumptive diagnosis of non-producing adrenal carcinoma was made. Laparoscopic of right adrenalectomy was carried out. Macroscopic appearance of the resected right adrenal surface showed a 3.0 × 2.5 cm whitish-grayish multicystic lesion with solid areas that replaced all adrenal medulla and almost all the cortex. Microscopic examination revealed multiple cystic channels lined with endothelial cells without atypia or mitotic figures. On immunohistochemical examination, anti-calretinin, anti-D2-40 and anti-Factor VIII were all positive confirming the diagnosis of adrenal cystic lymphangioma. It is a rare pathology that should be considered as part of the differential diagnosis during the evaluation of adrenal gland masses.

Many animals use their olfactory systems to learn to avoid dangers, but how neural circuits encode naive and learned olfactory preferences, and switch between those preferences, is poorly understood. Here, we map an olfactory network, from sensory input to motor output, which regulates the learned olfactory aversion of Caenorhabditis elegans for the smell of pathogenic bacteria. Naive animals prefer smells of pathogens but animals trained with pathogens lose this attraction. We find that two different neural circuits subserve these preferences, with one required for the naive preference and the other specifically for the learned preference. Calcium imaging and behavioral analysis reveal that the naive preference reflects the direct transduction of the activity of olfactory sensory neurons into motor response, whereas the learned preference involves modulations to signal transduction to downstream neurons to alter motor response. Thus, two different neural circuits regulate a behavioral switch between naive and learned olfactory preferences., Organismic and Evolutionary Biology, Physics

The Orthoflavivirus ilheusense (ILHV) is an arbovirus that was first isolated in Brazil in 1944 during an epidemiologic investigation of yellow fever. Is a member of the Flaviviridae family and it belongs to the antigenic complex of the Ntaya virus group. Psorophora ferox is the primary vector of ILHV and this study presents the isolation and phylogenetic analysis of ILHV in a pool of Ps. ferox collected in the state of Goiás in 2021. Viral isolation tests were performed on Vero cells and C6/36 clones. The indirect immunofluorescence test (IFI) was used to confirm the positivity of the sample. The positive sample underwent RT-qPCR, sequencing, and phylogenetic analysis. This is the first report of ILHV circulation in this municipality and presented close relationship between this isolate and another ILHV isolate collected in the city of Belém (PA)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The early detection and diagnosis of deaths in free-ranging non-human primates (NHPs) are key points for the surveillance of Yellow Fever (YF) in Brazil. The histopathological identification of infectious diseases remains very useful and reliable in the screening and detection of emerging zoonotic diseases such as YF. We surveyed data records and liver slides stained with hematoxylin and eosin from the Epizootics Surveillance Network to control YF, Ministry of Health of Brazil, to evaluate histopathological hallmarks for the diagnosis of the YF virus infection. We selected natural fatal cases in NHPs from the genera Alouatta spp., Callithrix spp., and Sapajus spp. with a positive immunohistochemical assay for YF in liver samples. Our findings showed the full-spectrum YF-associated hepatic lesions in all NHPs, but some histopathological findings differed in the distribution and intensity between the three genera. In our study, South American NHPs showed significant differences in the YF-associated hepatic histopathological features compared to fatal cases reported in humans., (Copyright © 2022 Elsevier B.V. All rights reserved.)

We present a 44-year-old man with multiple flat papules over a tattoo, diagnosed with acquired verruciform epidermodysplasia (EV). The lesions completely disappeared after 3 sessions of photodynamic therapy (PDT) with topical methyl aminolevulinate. PDT could be considered a treatment of choice in human papillomavirus lesions located over tattoos since it resolves the lesions while preserving the integrity of the design., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Accurate classification of postsurgical remission, and early recognition of recurrence are crucial to timely treat and prevent excess mortality in Cushing's Disease, yet the criteria used to define remission are variable and there is no consensus to define recurrence. Remission is defined as postsurgical hypocortisolemia, but delayed remission may occur. Recurrence is the return of clinical manifestations with biochemical evidence of hypercortisolism. The proper combination of tests and their timing are controversial. Reliable predicting tools may lead to earlier diagnosis upon recurrence. Many factors have been studied independently for prediction with variable performance. Novel artificial intelligence approaches seek to integrate these variables into risk calculators and machine-learning algorithms with an acceptable short-term predictive performance but lack longer-term accuracy. Prospective studies using these approaches are needed. This review summarizes the evidence behind the definitions of remission and recurrence and provide an overview of the available tools to predict and/or diagnose them., Competing Interests: Declaration of competing interest The authors report not to have conflict of interest concerning this manuscript., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Tattoo complications are an increasing reason for dermatological consultation in recent years, becoming a therapeutic challenge in some cases. We present a case of a suppurative granulomatous reaction in a tattoo resistant to oral antibiotic and topical corticosteroid treatment that resolved after four sessions of photodynamic therapy (PDT) with methyl aminolevulinate, possibly due to its anti-inflammatory and antimicrobial action, with an excellent aesthetic result. We suggest that the clinical utility of PDT may be expanded to treatment of tattoo complications, as well as in cutaneous infections of unknown microorganisms resistant to antibiotics., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS) has been shown to be an excellent technique for the discrimination of glass originating from different sources and for the association of glass originating from the same source. Typically, a match criterion is used to compare the elemental profile of the known sample to a questioned sample and if the glass samples are determined to "match" this may be followed by the use of a verbal scale to report the forensic practitioner's conclusion. This approach has several disadvantages: a fixed match criterion suffers from the "fall-off-the-cliff effect," the rarity of an elemental profile is not taken into account, and the use of a verbal scale to assign a weight of evidence may be considered as subjective and can vary by examiner. An alternative approach includes the use of a continuous likelihood ratio that provides a quantitative measure of the value of the evidence in support of any hypothesis and accounts for the rarity of an elemental profile through the use of a glass database. In the present study, two glass databases were used to evaluate the performance of the likelihood ratio; the first database includes 420 automotive windshield samples, while the second database includes 385 glass samples from casework. The multivariate kernel model was used for the calculation of the likelihood ratio. However, this model led to unreasonably large (or small) likelihood ratios. Thus, a calibration step, using the Pool Adjacent Violators (PAV) algorithm, was necessary in order to limit the likelihood ratio to reasonable values. The calibrated likelihood ratio presented rates of misleading evidence of <1.5% (for LRs<1 when objects came from the same source), and of <1.0% (for LRs>1 when objects came from different sources), which improved over the analogous ASTM false inclusion and false exclusion rates previously reported. In addition, the likelihood ratio limited the magnitude of the misleading evidence, providing only weak to moderate support for the incorrect hypothesis. Finally, most of the pairs found to present LR>1 when objects originated from different sources were explained by similarity of manufacturer of the glass source., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The cyst-forming protozoan parasite Neospora caninum is one of the main causes of bovine abortion worldwide and is of great economic importance in the cattle industry. Recent studies have revealed extensive genetic variation among N. caninum isolates based on multilocus microsatellite genotyping. Currently, the most extensive study reported is based on the N. caninum genotyping of 96 samples from four countries on two continents (Spain, Argentina, Germany and Scotland) that demonstrate different clusters of multilocus genotypes (MLGs) implicated in cattle abortions as well as the population sub-structuring of N. caninum, which is partially associated with the geographical origin. The aim of this study was to genotype N. caninum from aborted bovine foetuses that originated from Mexico within the region of Aguascalientes and to investigate their genetic diversity. Parasite DNA was detected in 27 out of the 63 analysed foetuses recovered from 10 different herds. Complete or nearly complete profiles based on 9 microsatellite markers were obtained from 11 samples. Diverse N. caninum MLGs were implicated in the occurrence of abortion in each herd. All of the Mexican MLGs differed from the MLGs previously determined for the Argentinean, Spanish, German and Scottish N. caninum populations. The Mexican MLGs failed to cluster by eBURST analyses. The MLG relationships using PCoA showed a close genetic relationship between the Spanish population and a portion of the Mexican population, but a more distant genetic relationship with the Argentinean genotypes. These results demonstrate the genetic diversity of N. caninum in the studied areas that differed from other populations of N. caninum around the world., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Synovitis, acne, pustulosis, hyperostosis, osteitis (SAPHO) syndrome represents the rare co-occurrence of sterile inflammatory osteoarticular disease in association with a variety of cutaneous manifestations. Oral involvement is uncommon. The etiology of SAPHO is complex and is likely the combined result of infectious, genetic, and immunologic factors. Due to diverse clinical presentations, SAPHO is difficult to diagnose. Here, we describe the case of a 74-year-old man, who had a history of SAPHO syndrome and presented with gingival pustules and sterile diffuse sclerosing osteomyelitis of the mandible. This is the first case report describing neutrophilic mucositis as a feature of SAPHO., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: In familial combined hyperlipidemia (FCHL) the severity of the dyslipidemia is determined by an overproduction of VLDL (very low density lipoprotein) particles and by its abnormal lipid composition. However, few are known regarding the metabolic factors that determine these abnormalities. We investigated the impact of metabolic factors on the number of atherogenic particles (apolipoprotein B level (apoB)) and the triglyceride content of very low-density lipoproteins (VLDLs-TG)., Methods: A cross-sectional study done in FCHL subjects and gender and age-matched healthy subjects. A clinical assessment, lipid profile and plasma concentrations of insulin, apolipoprotein CIII (apo CIII), apolipoprotein AII (apo AII), high sensitive C-reactive protein (HS-CRP), adiponectin and leptin were documented in 147 FCHL patients and 147 age-matched healthy subjects. Multivariate regression models were performed to investigate the independent determinants of VLDL-TG and apo B levels adjusting for confounding factors., Results: The variables that determined the VLDL-triglyceride content as a surrogate of VLDL composition were apo CIII (β=0.365, p<0.001), insulin (β=0.281, p<0.001), Apo AII (β=0.145, p<0.035), and adiponectin levels (β=-0.255, p<0.001). This model explained 34% of VLDL composition (VLDL-TG) variability. However, none of these variables were independent contributors of apo B-containing particles., Conclusions: In patients with FCHL apo CIII, apo AII and adiponectin are major novel factors determining the VLDL particle composition. However, such factors do not explain apo B-containing particles., (Copyright © 2014 Elsevier B.V. All rights reserved.)

This paper presents the possibilities of applying the likelihood ratio (LR) approach for the comparison problem to the data collected as a result of the Isotope Ratio Mass Spectrometry (IRMS) analysis targeted at lead (Pb)-isotope ratios. The assessment of the applied LR models performance was conducted by an Empirical Cross Entropy (ECE) approach. 35 glass samples were subjected to IRMS analysis and were described by Pb-isotope ratios: (208)Pb/(204)Pb, (207)Pb/(204)Pb, (206)Pb/(204)Pb, (208)Pb/(206)Pb, and (207)Pb/(206)Pb. Univariate and bivariate LR computations were performed, assuming normally distributed data subjected or not to a logarithmic transformation. Principal Component Analysis (PCA) was employed for creating orthogonal variables to propose an alternative LR model. It was found that the application of variable (208)Pb/(204)Pb seems to be promising as it delivers one of the lowest percentages of false positive and false negative rates as well as being the only variable for which an ECE plot gave satisfactory results., (Copyright © 2012 Elsevier B.V. All rights reserved.)

In this work, a selection of the best features for multivariate forensic glass classification using Scanning Electron Microscopy coupled with an Energy Dispersive X-ray spectrometer (SEM-EDX) has been performed. This has been motivated by the fact that the databases available for forensic glass classification are sparse nowadays, and the acquisition of SEM-EDX data is both costly and time-consuming for forensic laboratories. The database used for this work consists of 278 glass objects for which 7 variables, based on their elemental compositions obtained with SEM-EDX, are available. Two categories are considered for the classification task, namely containers and car/building windows, both of them typical in forensic casework. A multivariate model is proposed for the computation of the likelihood ratios. The feature selection process is carried out by means of an exhaustive search, with an Empirical Cross-Entropy (ECE) objective function. The ECE metric takes into account not only the discriminating power of the model in use, but also its calibration, which indicates whether or not the likelihood ratios are interpretable in a probabilistic way. Thus, the proposed model is applied to all the 63 possible univariate, bivariate and trivariate combinations taken from the 7 variables in the database, and its performance is ranked by its ECE. Results show remarkable accuracy of the best variables selected following the proposed procedure for the task of classifying glass fragments into windows (from cars or buildings) or containers, obtaining high (almost perfect) discriminating power and good calibration. This allows the proposed models to be used in casework. We also present an in-depth analysis which reveals the benefits of the proposed ECE metric as an assessment tool for classification models based on likelihood ratios., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Background: Given the elevated potential for primary or transmitted drug resistance (TDR) among newly HIV-infected individuals, there is a need for a deeper understanding of the baseline resistance patterns present in young men of color who have sex with men., Methods: Genotypic data were collected for participants aged 13-24 who were enrolled from seven sites. Univariate and bivariate methods were used to describe the prevalence of TDR and characteristics associated with TDR., Results: Of the 296 individuals participating in the substudy, 145 (49%) had baseline genotypes. The majority of the individuals were African American (65%) and gay-identified (70%). There was significant variation in genotype availability by site (p < .001). Major surveillance drug resistance mutations were present in 28 subjects (19.3%); the majority were non-nucleoside reverse transcriptase inhibitor mutations (12.4%). Subjects with TDR were less likely to have used alcohol on 1 or more days in the prior 2 weeks. Location was not associated with acquisition of TDR., Conclusions: There was a high rate of TDR in a geographically and racially diverse sample of HIV-infected young men of color who have sex with men. This represents a serious public health concern given the young age of this sample and the potential need for long-term antiretroviral therapy. These findings underscore the critical roles of both early case identification and secondary prevention., (Copyright © 2011 Society for Adolescent Health and Medicine. All rights reserved.)

The nervous system consists of hundreds of billions of neurons interconnected into the functional neural networks that underlie behaviors. The capacity of a neuron to innervate and function within a network is mediated via specialized cell junctions known as synapses. Synapses are macromolecular structures that regulate intercellular communication in the nervous system, and are the main gatekeepers of information flow within neural networks. Where and when synapses form determines the connectivity and functionality of neural networks. Therefore, our knowledge of how synapse formation is regulated is critical to our understanding of the nervous system and how it goes awry in neurological disorders. Synapse formation involves pairing of the pre- and postsynaptic partners at a specific neurospatial coordinate. The specificity of synapse formation requires the precise execution of multiple developmental events, including cell fate specification, cell migration, axon guidance, dendritic growth, synaptic target selection, and synaptogenesis (Juttner and Rathjen in Cell. Mol. Life Sci. 62:2811, 2005; Salie et al., in Neuron 45:189, 2005; Waites et al., in Annu. Rev. Neurosci. 28:251, 2005). Remarkably, during the development of the vertebrate nervous system, these developmental processes occur almost simultaneously in billions of neurons, resulting in the formation of trillions of synapses. How this remarkable specificity is orchestrated during development is one of the outstanding questions in the field of neurobiology, and the focus of discussion of this chapter. We center the discussion of this chapter on the early developmental events that orchestrate the process of synaptogenesis prior to activity-dependent mechanisms. We have therefore limited the discussion of important activity-dependent synaptogenic events, which are discussed in other chapters of this book. Moreover, our discussion is biased toward lessons we have learned from invertebrate systems, in particular from C. elegans and Drosophila. We did so to complement the discussions from other chapters in this book, which focus on the important findings that have recently emerged from the vertebrate literature. The chapter begins with a brief history of the field of synaptic biology. This serves as a backdrop to introduce some of the historically outstanding questions of synaptic development that have eluded us during the past century, and which are the focus of this review. We then discuss some general features of synaptic structure as it relates to its function. In particular, we will highlight evolutionarily conserved traits shared by all synaptic structures, and how these features have helped optimize these ancient cellular junctions for interneural communication. We then discuss the regulatory signals that orchestrate the precise assembly of these conserved macromolecular structures. This discussion will be framed in the context of the neurodevelopmental process. Specifically, much of our discussion will focus on how the seemingly disparate developmental processes are intimately linked at a molecular level, and how this relationship might be crucial in the developmental orchestration of circuit assembly. We hope that the discussion of the multifunctional cues that direct circuit development provides a conceptual framework into understanding how, with a limited set of signaling molecules, precise neural wiring can be coordinated between synaptic partners.

Myeloperoxidase (MPO) is a dominating enzyme of circulating polymorphonuclear neutrophils that catalyzes the two-electron oxidation of chloride, thereby producing the strong halogenating agent hypochlorous acid (ClO(-)/HOCl). In absence of MPO the tripeptide Pro-Gly-Gly reacts with HOCl faster than the amino acid taurine (2-aminoethanesulfonic acid, Tau), while the MPO-mediated chlorination shows reverse order. A comparative study of the enzymatic oxidation of both substrates at pH 4.0-6.0, varying H2O2 concentration is presented. Initial and equilibrium rates studies have been carried on, reaction rates in the latter being slower due to the chemical equilibrium between MPO-I and MPO-II-HO2. A maximum of chlorination rate is observed for Pro-Gly-Gly and Tau when [H2O2] approximately 0.3-0.7 mM and pH approximately 4.5-5.0. Several mechanistic possibilities are considered, the proposed one implies that chlorination takes place via two pathways. One, for bulkier substrates, involves chlorination by free HOCl outside the heme cavity; ClO(-) is released from the active center, diffuses away the heme cavity, and undergoes protonation to HOCl. The other implies the existence of compound I-Cl(-) complex (MPO-I-Cl), capable of chlorinating smaller substrates in the heme pocket. Electronic structure calculations show the size of Pro-Gly-Gly comparable to the available gap in the substrate channel, this tripeptide being unable to reach the active site, and its chlorination is only possible by free HOCl outside the enzyme.

An experimental and theoretical investigation on crystal forms of the popular and ubiquitous pharmaceutical Minoxidil is presented here. A new crystallization method is presented for Minoxidil (6-(1-piperidinyl)-2,4-pyrimidinediamide 3-oxide) in ethanol-poly(ethylene glycol), yielding crystals with good quality. The crystal structure is determined at low temperature, with a final R value of 0.035, corresponding to space group P2(1) (monoclinic) with cell dimensions a = 9.357(1) A, b = 8.231(1) A, c = 12.931(2) A, and beta = 90.353(4) degrees . Theoretical calculations of the molecular structure of Minoxidil are set forward using empirical force fields and quantum-mechanical methods. A theoretical prediction for Minoxidil crystal structure shows many possible polymorphs. The predicted crystal structures are compared with X-ray experimental data obtained in our laboratory, and the experimental crystal form is found to be one of the lowest energy polymorphs.

Myeloperoxidase (MPO) catalyzes the two-electron oxidation of chloride, thereby producing hypochlorous acid (HOCl). Taurine (2-aminoethane-sulfonic acid, Tau) is thought to act as a trap of HOCl forming the long-lived oxidant monochlorotaurine [(N-Cl)-Tau], which participates in pathogen defense. Here, we amend and extend previous studies by following initial and equilibrium rate of formation of (N-Cl)-Tau mediated by MPO at pH 4.0-7.0, varying H(2)O(2) concentration. Initial rate studies show no saturation of the active site under assay conditions (i.e. [H(2)O(2)] > or = 2000 [MPO]). Deceleration of Tau chlorination under equilibrium is quantitatively described by the redox equilibrium established by H(2)O(2)-mediated reduction of compound I to compound II. At equilibrium regime the maximum chlorination rate is obtained at [H(2)O(2)] and pH values around 0.4mM and pH 5. The proposed mechanism includes known acid-base and binding equilibria taking place at the working conditions. Kinetic data ruled out the currently accepted mechanism in which a proton participates in the molecular step (MPO-I+Cl(-)) leading to the formation of the chlorinating agent. Results support the formation of a chlorinating compound I-Cl(-) complex (MPO-I-Cl) and/or of ClO(-), through the former or even independently of it. ClO(-) diffuses away and rapidly protonates to HOCl outside the heme pocket. Smaller substrates will be chlorinated inside the enzyme by MPO-I-Cl and outside by HOCl, whereas bulkier ones can only react with the latter.

Beta-6 Integrin, tenascin-C, and MMP-1 (matrix metalloproteinase-1) are invasion-related proteins that are frequently overexpressed in many human malignancies. The objective of this study was to determine whether there is overexpression of these molecules in three types of salivary neoplasms showing markedly different behavior. A total of 55 formalin-fixed, paraffin-embedded archived specimens comprising 19 adenoid cystic carcinomas (ACC), 18 polymorphous low-grade adenocarcinomas (PLGA) and 18 pleomorphic adenomas (PA) were utilized in this study. A standard immunohistochemical technique was used to determine the expression levels of beta-6 integrin, tenascin-C, and matrix metalloproteinase-1 (MMP-1) proteins. Sections were assessed semiquantitatively, and tumors were divided into two groups, low-expressors (0-1+) and high-expressors (2-3+) for statistical analysis. Staining was graded as 0 (<1% positive tumor cells), 1+ (<25% positive tumor cells), 2+ (25-50% positive tumor cells), and 3+ (>50% positive cells). The results showed that the malignant tumors were higher expressors of beta-6 than the benign tumors. ACCs showed significantly higher expression of beta-6 than PAs (p=0.04). No significant difference was observed between ACCs and PLGAs. beta-6 expression was rarely seen in normal salivary gland epithelium and was occasionally present in mucosa overlying the tumors. PAs were high-expressors of tenascin-C with a significant difference relative to ACCs (p=0.03). A majority of tumors in all three tumor types showed high expression of MMP1 with expression significantly greater in the PAs compared to ACCs (p=0.008). We conclude that ACCs and PLGAs express beta-6, tenascin-C, and MMP-1, but that their expression patterns are not significantly different. beta-6 appears to be more closely associated with the malignant tumors, and MMP-1 more closely associated with the benign tumors. We believe that beta-6, tenascin-C, and MMP-1 proteins are part of the molecular repertoire used by salivary tumors for malignant invasion and benign tumor expansion.

In this study we now show that deposition of the mesenchymal matrix marker, tenascin-C (TN-C), is mediated through beta3 expression and activation of Src. There was a striking upregulation of TN-C matrix organization in cell lines expressing beta3 and activated Src when compared to cell lines with neither of these attributes. When beta3 function was suppressed so was the deposition of TN-C. The same was true for function and activation of Src. When Src was inactive, the deposition of TN-C was low. We also determined that one of the downstream effectors of Src, MAPK, was also required to promote TN-C deposition. When MAPK activation was inhibited, TN-C deposition was also decreased. MMP activation is also implicated in TN-C deposition. The broad spectrum MMP inhibitor, GM6001, suppressed TN-C organization. These results indicate that beta3 integrin ligand binding and the activation of the Src/MAPK/MMP pathway modulate deposition of TN-C.

The integrin beta6 has been shown to promote invasion and experimental metastasis by oral squamous cell carcinoma (SCC). In this study, we demonstrate that the expression of beta6 by oral SCC9 cells increased activation of the UPA --> MMP3 --> MMP9 pathway. We also demonstrate that the deposition of fibronectin and tenascin-C matrices by SCC9beta6 cells and peritumor fibroblast cocultures is counter-regulated by the UPA --> MMP3 --> MMP9 pathway. Suppression of individual components of this pathway increased the deposition of fibronectin, but decreased tenascin-C matrix assembly by the cocultures. When the SCC9beta6/PTF cocultures were incubated with TGFbeta1, the deposition of fibronectin and tenascin-C as well as the activation of MMP3 and MMP9 was increased. These results indicate that MMP3, MMP9, and TGFbeta1 are important for the modulation, composition, and maintenance of the ECM in oral SCC.

Expression of beta 3 integrins is increased in invasive melanoma. In this study we show that K1735 cell proliferation is enhanced by the expression of either beta 3 or a constitutively active Src. We investigated possible modulators of FN matrix assembly and found that matrix metalloproteinase 2 (MMP2) was activated by alpha v beta 3. alpha v beta 3 integrin was localized to focal contacts whereas alpha v beta 5 was peripherally distributed. MMP2 was also activated by expression of CASrc. MMP2 activation inversely correlated with FN matrix assembly, in that it dramatically reduced the organization of a FN matrix. K1735 cell migration on VN and invasion through a reconstituted basement membrane were decreased in the presence of anti-MMP2 antibodies. These results demonstrate that the expression of the alpha v beta 3 complex modulates melanoma cell behavior including activation of Src, organization of the cytoskeleton, assembly of the extracellular matrix, cell motility, and activation of MMP2.

Objective: Oral warts arising in human immunodeficiency virus (HIV) infection occasionally show marked epithelial dysplasia. However, anecdotal evidence suggests that they do not progress to oral squamous cell carcinoma (SCC). Therefore, we evaluated lesions for expression of proteins (tenascin-C, beta6 integrin, and matrix metalloproteinase-1[MMP1]) that have been identified as important in the invasive phase of oral SCC., Study Design: Twenty-two oral dysplastic warts from 22 patients and 5 oral SCCs were stained for human papillomavirus (HPV) antigen, proliferation protein Ki-67, tenascin-C, beta6, and MMP1 by immunohistochemical methods. For comparison, 5 nondysplastic warts each from HIV-positive and HIV-negative patients and 5 normal mucosa specimens were included. Sections were semiquantitatively assessed, and results were compared. Because MMP1 was the lowest or least expressed interface protein, MMP1 mRNA was quantitatively assessed from formalin-fixed paraffin-embedded tissue in selected cases with quantitative reverse transcription-polymerase chain reaction., Results: Twenty of 22 dysplastic warts stained positive for human papillomavirus common antigen, and all warts showed high proliferative fractions similar to SCCs. Tenascin-C and beta6 were variably expressed by the dysplastic warts but were consistently expressed at high levels in the SCCs. MMP1 protein levels were negative or low in 20 of 22 in dysplastic warts, but were elevated in 4 of 5 SCCs. MMP1 mRNA analysis indicated that message was low in 4 dysplastic warts and also suggested that protein translation was incomplete in 3 of the warts., Conclusion: We conclude that invasion-associated proteins are underexpressed in oral dysplastic warts in HIV-positive men. However, until these patients are followed for extended periods, the risk of development of SCC from oral dysplastic warts remains unknown.

Floor of the mouth squamous cell carcinomas exhibit many characteristics that suggest they represent a distinct biological subset within head and neck tumors. The features of preinvasive lateral intraepithelial spread, high rate of conversion of intraepithelial neoplasia to invasive carcinoma, and high incidence of occult metastases, suggest the importance of motility-associated proteins in the pathogenesis of these lesions. Two such proteins, tenascin and beta 6 integrin, are generally overexpressed in squamous carcinomas, and may play a central role in the invasive process of floor of the mouth lesions. The purpose of this study was to evaluate in situ and invasive squamous cell carcinomas from the floor of the mouth for the expression of tenascin and beta 6 integrin. Twenty lesions each of floor of the mouth in situ carcinomas and squamous cell carcinomas, and 10 normal controls were stained for tenascin and beta 6 using a standard immunohistochemical protocol for formalin-fixed specimens. Sections were assessed for staining intensity, pattern, and co-localization. Tenascin was highly expressed at the keratinocyte-connective tissue interface of both in situ and invasive carcinomas. beta 6 was expressed in basal keratinocytes of all in situ and invasive lesions, but was not evident in any of the control epithelia. There was no significant difference in staining of in situ and invasive carcinomas, but there was a significant difference in staining between these lesions and controls. Staining was colocalized in serial sections, supporting a receptor-ligand relationship. Both tenascin and beta 6 were weakly expressed in dysplastic areas adjacent to carcinomas suggesting that changes in the expression of these proteins occurs prior to the invasive phenotype. We conclude that tenascin and beta 6 are overexpressed in in situ and invasive floor of the mouth carcinomas, but that transgression of the basement membrane by neoplastic epithelial cells requires additional changes to the keratinocyte molecular profile.

Oral squamous cell carcinoma (SCC) is characterized by invasive growth and the propensity for distant metastasis. The expression of specific adhesion receptors promotes defined interactions with the specific components found within the extracellular matrix (ECM). We previously showed that the alpha v beta 6 fibronectin receptor is highly expressed in oral SCC. Here we forced expression of the beta 6 subunit into poorly invasive SCC9 cells to establish the SCC9 beta 6 cell line and compared these two cell lines in several independent assays. Whereas adhesion to fibronectin was unaffected by the expression of beta 6, migration on fibronectin and invasion through a reconstituted basement membrane (RBM) were both increased. Function-blocking antibodies to alpha v beta 6 (10D5) reduced both migration on fibronectin and invasion through an RBM, whereas anti-alpha 5 antibodies were effective only in suppressing migration on fibronectin, not invasion. Expression of beta 6 also promoted tumor growth and invasion in vivo and modulated fibronectin matrix deposition. When grown as a co-culture with SCC9 cells, peritumor fibroblasts (PTF) organized a dense fibronectin matrix. However, fibronectin matrix assembly was decreased in co-cultures of SCC9 beta 6 cells and PTF and this decrease was reversed by the addition of function-blocking anti-alpha v beta 6 antibodies. The expression of beta 6 also resulted in increased levels of matrix metalloproteinase 3. Addition of the general MMP inhibitor GM6001 to SCC9 beta 6/PTF co-cultures dramatically increased fibronectin matrix assembly in a similar fashion as incubation with anti-alpha v beta 6 antibodies. These results demonstrate that expression of beta 6 (1) increases oral SCC cell motility and growth in vitro and in vivo; (2) negatively affects fibronectin matrix assembly; and (3) stimulates the expression and activation of MMP3. We suggest that the integrin alpha v beta 6 is a key component of oral SCC invasion and metastasis through modulation of MMP-3 activity.