Microm Thick Sections Research Articles

No implication of Simian virus 40 in pathogenesis of malignant pleural mesothelioma in Slovenia.

Malignant mesothelioma is predominantly caused by asbestos exposure, although the association of Simian virus 40 in its pathogenesis is currently still under debate. Simian virus 40, a DNA rhesus monkey virus with oncogenic properties, accidentally contaminated early batches of polio vaccine in the 1960s. In the 1990s, viral sequences and proteins were discovered in several human tumors, which triggered research to find a link between Simian virus 40 and human cancers, especially malignant mesothelioma. The aim of our study was to establish an effective laboratory procedure for Simian virus 40 detection and to investigate the presence of Simian virus 40 DNA and small t antigen in mesothelioma samples from Slovenian patients. Paraffin-embedded malignant pleural mesothelioma specimens from 103 Slovenian patients were collected and used for total DNA isolation and real-time polymerase chain reaction for Simian virus 40 small t and large T DNA analysis. Special attention was devoted to primer design, good laboratory practice and polymerase chain reaction contamination prevention. Polymerase chain reaction products were sequenced and BLAST aligned. One 5 microm thick paraffin section from each patient's tissue block was stained with hematoxylin and eosin for histological typing and one for immunohistochemical detection of Simian virus 40 small t antigen using a monoclonal antibody against Simian virus 40 (Pab280). SV40-expressing Wi-38 cells were used as positive control in both PCR and immunohistochemistry. In real-time polymerase chain reaction analyses, only 4 samples gave products with primer pairs amplifying small t antigen and were inconsistent and poorly reproducible. BLAST alignment showed no homology with any deposited SV40 sequences. No immunopositive staining for SV40 small t antigen was found in any of the samples. We found no evidence of SV40 presence in tissue samples from 103 Slovenian patients with malignant pleural mesothelioma. Asbestos exposure remains the main risk factor for malignant pleural mesothelioma in Slovenia.

Spherulites of Amyloid-β42 In Vitro and in Alzheimer's Disease

Several amyloidogenic proteins including insulin, beta-lactoglobulin, and albumin form spherulites in vitro under non-physiological conditions. These micrometer-sized, roughly spherical structures are composed of ordered arrays of amyloid fibrils in radial arrangements which, characteristically, show a typical Maltese cross pattern of light extinction under the polarizing microscope. The physiological significance of amyloid spherulites is unknown though in Alzheimer's disease, senile plaques composed primarily of beta sheets of amyloid-beta (Abeta)42 have, very occasionally, been shown to give a Maltese cross pattern of light extinction under crossed polarizers. Herein we describe the first observation of the formation in vitro of spherulites of Abeta42. They were formed under near-physiological conditions in which the beta sheet conformation of pre-formed aggregates of Abeta42 had been abolished following the addition of an excess of copper. Incubation of these preparations at 37 degrees C for up to 9 months resulted in the formation of globular structures, 5-20 microm in diameter, which exhibited a Maltese cross pattern of light extinction typical of spherulites. Near-identical spherulitic structures were also observed in abundance in 30 microm thick sections of Alzheimer's disease brain tissue. Synchrotron x-ray fluorescence showed that the location of these spherulites in AD tissue coincided with locally elevated concentrations of tissue copper. The formation in vitro of spherulites of Abeta42 which morphologically appeared analogous to spherulitic structures observed in vivo strongly supports the hypothesis that spherulites and senile plaques in AD tissue are one and the same structures and that their ultimate formation may involve copper.

Anatomía microscópica y metabolitos secundarios volátiles en tres estadios del desarrollo de las inflorescencias de Lantana camara (Verbenaceae)

Plants of the Verbenaceae family, like L. camara, have called the attention of researchers, not only because of its high diversity and its distribution around the world, but also for its variable use as popular medicine to treat diseases like tetanus, rheumatism and malaria, and as bactericide and insecticide. To assess this, the morphology and ontogeny of the inflorescences of Lantana camara and the chemical composition of volatile secondary metabolites were analyzed at three different ontogeny stages. Plants were collected from the experimental crop area in CENIVAM, Bucaramanga, Colombia. Fresh inflorescence stages were established and analyzed using a stereoscopic microscope, fixed in FAA and included in parafine. Transversal and longitudinal 10 microm thick sections were prepared using a rotative microtome, safranine-fastgreen stained and were observed and photographed using a light microscope. The chemical composition of volatile secondary metabolites were analyzed for each stage. The analytes, obtained from 0.7 g of plant, were isolated by solid phase micro-extraction in the headspace mode (HS-SPME) and were placed in 20 ml vials. The components were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Stage I was microscopically characterized by an immature development in which the meristematic differentiation begins with a mass of cells. In Stage II, the morphogenetic movement gives way to the formation of the respective floral sexual structures, calyx and corolla. In Stage III, the different organs are conspicuous: four stamens epipetals and didynamous, monocarpelar, biloculate and globose gynoecium, upper ovary and lateral stigma; the flowers are hermaphroditic. The main secondary metabolites detected by GC-MS were bicyclosesquiphellandrene, E-beta-farnesene, E-beta-caryophyllene, gamma-muurolene + gamma-curcumene and alpha-zingiberene. Nevertheless, this study reports for the first time in plant species alpha-gurjunene, gamma-amorphene, alpha-muurolene, sesquithujene, alpha-trans-bergamotene and trans-cadina-1,4-diene. The diversity of compounds found can be only explained by the extraction methods employed, the developmental stages and section of the plant, the geographic conditions, collection time and the genetic constitution of the evaluated species.

Non-fatal injuries to the cervical spine facet joints after a fatal motor vehicle crash

Injuries to the lower cervical spine facet joints may be identified in people killed in road traffic crashes. However, many of these injuries are not identified on diagnostic imaging procedures despite histological evidence of injury. This case report illustrates possible non-lethal pathological conditions in the facet joints following fatal road traffic trauma. The cervical spine segments C4 to C7 were removed en bloc and examined with magnetic resonance imaging, computed tomography and conventional radiology. The specimen was fixated in alcohol, embedded in methylmethacrylate, and divided into 3 mm thick parasaggital slices from where 10 microm thick sections were produced and evaluated with light microscopy. Injuries identified on microscopy included haemarthrosis, capsular avulsion, synovial fold contusions and subchondral fractures. None of the injuries were detected on diagnostic imaging or at routine autopsy. Discrete injuries to the cervical spine facet joints were identified using supplemental histological procedures in this case report. Post-mortem diagnostic imaging did not reveal these injuries and future studies should investigate the nature, prevalence and clinical relevance of such lesions.

Amyloidose in Leberbiopsien

We reassessed the histopathology and origin of amyloid in liver biopsies. All liver biopsies were retrieved from a series of 588 cases with histologically confirmed amyloidosis submitted between February 2006 and January 2009 to the Amyloid Registry of the Charité University Hospital. Liver biopsies had been fixed in formalin and embedded in paraffin. 3-5 microm thick paraffin sections were stained with hematoxylin and eosin and Congo red. Amyloid was classified immunohistochemically, using antibodies directed against amyloid P-component, AA amyloid, apolipoprotein AI, fibrinogen, lysozyme, lambda- and kappa-light chain, and transthyretin. Amyloid was found in 46 liver biopsies (29 men, 17 women; mean age 60 years, range 34-87 years). Immunohistochemical classification succeeded in 42 cases. AL amyloidosis was present in 40 (87%) cases and was further categorized into AL amyloid of lambda-light chain origin in 26 (57%) cases, and kappa-light chain origin in 14 (30%) cases. ATTR and AA amyloidosis were found in a single patient each (2%). In 4 (9%) cases, amyloid remained unclassified. Hepatic amyloidosis is most commonly AL amyloid of lambda- and kappa-light chain origin and is often associated with marked parenchymal atrophy.

Direct Sampling from Human Liver Tissue Cross Sections for Electrophoretic Analysis of Doxorubicin

After chemoembolization of the liver with doxorubicin (Dox), this drug and its metabolites are not homogeneously distributed in this organ. The distribution cannot be easily measured making it difficult to assess how the drug performs in different tissue regions. Here we report a technique for sampling tissue cross sections that can analyze the contents of micrometer size regions. The tissue cross sections were from the explanted liver of a hepatocellular carcinoma patient. Samples were directly aspirated from a 5 microm thick tissue cross section into a 50 microm i.d. capillary where the tissue was solubilized with a separation buffer containing sodium dodecyl sulfate. Upon sample dissolution, Dox and natively fluorescent compounds were separated and detected by micellar electrokinetic chromatography with laser-induced fluorescence detection. Sampling reproducibility and recovery were assessed using 10% (w/v) gelatin as tissue mimic. Sampling from gelatin slices containing Dox revealed a relative standard deviation of 13%, which was comparable to that of sampling from solution. Dox recovery was 82% +/- 16% (n = 5). When sampling tumor and nontumor tissue regions, samples could be taken from the same region 100 microm apart. Atomic force microscopy was used to determine that each sample was 8.4 +/- 1.0 pL in volume which made it possible to determine Dox concentrations in the ranges of 0.4-1.3 and 0.3-0.5 microM for the samples taken from tumor and nontumor regions, respectively. The results demonstrated the feasibility of sampling, detection, and quantification of Dox in micrometer size regions, which could be a useful resource for analyzing the Dox concentration and distribution in highly heterogeneous tissues.

Histología y morfometría del ojo del pez dulceacuícola Paracheirodon axelrodi (Characiformes: Characidae)

The visual system of Teleost fishes has a wide range of adaptations. The eye has suffered modifications, mainly towards better sensitivity and visual resolution according to specific environmental conditions. In that way, this organ is so important for survival. In spite of being the most commercialized Orinoquia species, and one of the most important ornamental fish worldwide, there are almost no studies on the communication systems of P. axelrodi (including vision). We studied the eye in ten individuals of P. axelrodi, with a mean total length of 2.7 cm (SD = 0.2, n = 20). We measured eye relative size (TRO) after sacrifice. Heads were processed for hematoxylin-eosin, and 8 microm thick sections were obtained. Images were digitalized for histological identification and morphometrics. The relation between TRO and body length was 9.8% (SD= 1.1, n= 20); lens was spherical with a relative size of 64.3% (SD = 4.9, n = 20). Rete mirabilis choroid was composed mainly by capillaries, while sclera was composed of cartilaginous tissue. Retina with eight layers and two membranes. An areae centrales was observed in the posterior retina. Iris covered by a simple cubic epithelium. Cornea of endotelial tissue with two membranes. Pupil almost circular, with a naso-caudal elongation. Measures of retina thickness were as follows: lateral retina 144.5 microm (SD = 5.4, n = 20), temporal retina 210.4 microm (SD = 14.8, n = 20). Lens measurements were: dorso-ventral 759.6 microm (SD = 31.7, n = 20), antero-posterior 763.4 microm (SD = 30.7, n = 20). Cornea thickness was 27.6 microm (SD = 4.3, n = 20); iris thickness was 18.9 microm (SD = 1.9, n = 20); rete mirabilis choroid 22.2 microm (SD = 3.2, n = 20) and sclera: 15.9 microm (SD = 1.2, n = 20). This species has a typical Teleost eye conformation. The retina was thicker in the posterior area, probably related to the presence of neuron groups. This modification allows for a better sharpness and visual resolution. The TRO, areae centrales in the retina, and thin cornea, suggests the need of a well developed visual system resulting from a predatory diet.

Skin fungal biocontamination and the skin hydrogel pad test

Previous observations have revealed that environmental nondermatophyte molds (NDM) can grow inside specific hydrogel pads (LaserAid). Some of these NDM might be responsible for superficial and invasive mycoses as well as for allergic respiratory and cutaneous disorders. The load of NDM propagules in the environment is considered to be an important risk factor for all these diseases. It is postulated that the quantification of the responsible fungi deposited at the skin surface may be an indicator of a recent exposure to environmental fungi. The aim of the present study was to assess using the LaserAid hydrogel pads, the density of living NDM adhering to the skin surface of healthy subjects. Sterile hydrogel pads were applied in a repeat procedure onto the normal-looking skin of the palms and face of 35 healthcare workers who were active in low exposure areas. Similar samplings were performed after washing the skin with a regular skin cleanser, or after applying an alcohol solution or a povidone iodine solution. As controls, 20 sterile pads were exposed for a few minutes to ambient air of the laboratory without any contact with the skin. Each of these samples was stored for 2 weeks at room temperature in a clean protected environment. After that period, visual inspection of the pads was followed by microscopic examination of PAS-stained 6 microm-thick sections. In addition, mycological cultures were performed from pieces of the pads deposited onto Sabouraud agar plates. While 19/20 air-exposed samples were not contaminated by environmental air-borne fungi, 61/70 of the initial skin samplings and 6/70 of the repeat skin samplings showed foci of fungal colonization confirmed by microscopic examination. No specific differences were disclosed between the face and palm samplings. Cultures revealed the presence of NDM in the majority (64/67) of the colonized pads, and a few Candida albicans contaminations (3/67) were also disclosed. The cleansing with a non-antimicrobial product as well as disinfecting procedures performed before sampling markedly decreased the mycoflora without, however, clearing the skin of NDM and yeasts. In conclusion, the hydrogel pad procedure brings information about potential environmental skin contamination by NDM and commensal yeasts. The regular cleansing and disinfecting procedures do not eradicate these fungi from the skin surface.

Calcification of intervertebral discs in the dachshund: a radiographic and histopathologic study of 20 dogs

BackgroundThe purpose of the study was to compare radiographic and histopathologic findings with regard to number and extent of calcified discs in the dachshund.MethodsThe intervertebral discs of 20 dachshunds were subjected to a radiographic and histopathologic examination. The dogs were selected randomly from clinical cases euthanased for reasons unrelated to research at the Norwegian School of Veterinary Science. Lateral radiographs were taken of the vertebral columns after removing them from the carcasses. The histopathologic examination included 5 μm thick sections in the transverse plane, stained with hematoxylin-eosin and von Kossa. Radiographs and histological sections were evaluated independently.ResultsA total of 148 (28.5%) calcified discs were identified at the radiographic and 230 (45.7%) at the histopathologic examination. Of 92 discs found to be calcified by histopathology, but not by radiography, the degree of calcification was evaluated as 'slight' in 84 (91.3%). All the intervertebral discs (n = 138) that were found to be calcified by radiography were also found to be calcified by histopathology.ConclusionA sensitivity of 0.6 and specificity of 1.0 for radiography was calculated when using histopathology as the gold standard.

Effects of child formula dentifrices on artificial caries like lesions using in vitro pH-cycling: preliminary results

To compare the remineralisation effects of different child dentifrices on primary teeth. In vitro single-section technique and pH-cycling model. Primary teeth were painted with nail varnish, leaving a 1 mm wide window before placing in demineralising solution for 96hr to produce artificial carious lesions 150-200 microm deep. Teeth were longitudinally cut into approximately 100-150 microm thick sections and assigned to three groups (n = 7). Sections in Group A were exposed to Perioe Children's Toothpaste (LG, Korea), Group B to Colgate Pokemon (Colgate-Palmolive, Thailand) and Group C to Vicco (Vicco Laboratories, India). Polarised light microscopy and microradiography was used to evaluate lesion depth, before and after 7 days pH cycle. Mean lesion depths in Groups A and C increased by 11% and 14% respectively, while Group B demonstrated a lesion reduction of 3%. Comparisons using ANOVA and Student-Newman-Keuls tests showed that Groups A and C were significantly different from Group B (p < 0.05), but there was no significant difference between Groups A and C. Based on the data obtained, Colgate Pokemon remineralised initial carious lesions. In addition, when compared to Colgate Pokemon, Perioe Children's Toothpaste failed to show 'healing' efficacy even though it is claimed to contain fluoride.

Effects of dentifrices on artificial caries like lesions: an in vitro pH cycling study

To evaluate remineralising effects of three dentifrices, Vicco by Vicco Labs, India, Perioe Cavity Care by LG Care, Korea and Colgate Total by Colgate-Palmolive, Thailand on artificial caries like lesions. In vitro single-section and pH cycling model. Extracted third molars were painted with nail varnish, leaving 1 mm wide windows before placing demineralising solution for 96h to produce artificial carious lesions 150-200 miocrom deep. Teeth were then cut longitudinally into 100-150 microm thick sections and randomly assigned to three groups. Polarised light microscopy and microradiography were used to evaluate lesion depth, before and after the 10 day pH cycle. Sections in Group A were treated with Vicco by Vicco Labs, India (-ve control), Group B were treated with Perioe Cavity Care by LG Care, Korea and Group C were treated with Colgate Total by Colgate-Palmolive, Thailand. Mean lesion depths in Groups B and C reduced by 7% and 12% respectively, whereas lesion depth increased by 13% in Group A. Comparisons made among groups using ANOVA and Student-Newman-Keuls tests showed that three groups were significantly different from each other (p < 0.05). Perioe Cavity Care and Colgate Total remineralised initial carious lesions. However, the remineralising potential of Colgate Total was higher than that of Perioe Cavity Care.

MALDI‐ion mobility‐TOFMS imaging of lipids in rat brain tissue

While maintaining anatomical integrity, matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) has allowed researchers to directly probe tissue, map the distribution of analytes and elucidate molecular structure with minimal preparation. MALDI-ion mobility (IM)-orthogonal time-of-flight mass spectrometry (oTOFMS) provides an advantage by initially separating different classes of biomolecules such as lipids, peptides, and nucleotides by their IM drift times prior to mass analysis. In the present work the distribution of phosphatidlycholine and cerebroside species was mapped from 16 microm thick coronal rat brain sections using MALDI-IM-oTOFMS. Furthermore, the use of gold nanoparticles as a matrix enables detection of cerebrosides, which although highly concentrated in brain tissue, are not easily observed as positive ions because of intense signals from lipids such as phosphatidlycholines and sphingomyelins.

Vagina vasorum dissection during D2 lymphadenectomy for gastric carcinoma

To explore the relationship between metastasis and vagina vasorum in the progress of gastric carcinoma and to find some facts and references for gastric surgeons. One hundred and seven specimens of left or right gastric arteries (55 left and 52 right) were gathered from 59 patients undergoing radical gastrectomy for gastric carcinoma. All the frozen specimens were cut into 3 microm-thick sections and stained with hematoxylin-eosin (HE) and immunohistochemical method separately. Cytokeratin (CK) and mesothelial cells (MC) were stained with immunohistochemical method. Cancer cells inside vagina vasorum were detected and the structure of artery wall was observed under microscope. Metastatic cancer cells or tubercles were found inside vagina vasorum in some stage III or IV specimens, but not in stage I or II specimens. Tumor cells in vagina vasorum were CK positive in 26 specimens of 14 tumors. Among them, stage III was found in 4 specimens of 2 tumors, and stage IV in 22 specimens of 12 tumors. None of these specimens was positive for MC. The positive rate of CK increased with TNM staging. Compared with the lower part, tumors in the upper and middle parts of stomach were more likely to metastasize into vagina vasorum. Vagina vasorum dissection should be performed during D2 lymphadenectomy for TNM stage III or IV gastric carcinoma.

Application of highly sensitive fluorescent dyes (CyDye DIGE Fluor saturation dyes) to laser microdissection and two-dimensional difference gel electrophoresis (2D-DIGE) for cancer proteomics

Proteome data combined with histopathological information provides important, novel clues for understanding cancer biology and reveals candidates for tumor markers and therapeutic targets. We have established an application of a highly sensitive fluorescent dye (CyDye DIGE Fluor saturation dye), developed for two-dimensional difference gel electrophoresis (2D-DIGE), to the labeling of proteins extracted from laser microdissected tissues. The use of the dye dramatically decreases the protein amount and, in turn, the number of cells required for 2D-DIGE; the cells obtained from a 1 mm2 area of an 8-12 microm thick tissue section generate up to 5,000 protein spots in a large-format 2D gel. This protocol allows the execution of large-scale proteomics in a more efficient, accurate and reproducible way. The protocol can be used to examine a single sample in 5 d or to examine hundreds of samples in large-scale proteomics.

A clinical and immunohistochemical study on gastrointestinal mesenchymal tumor (GIMT): RCAS1 as a predictor for recurrence of GIMT

Gastrointestinal mesenchymal tumors (GIMTs) are the most common mesenchymal tumors of the gastrointestinal tract. RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is a cancer cell-surface antigen and has been identified as a prognostic factor in several cancer types. It is thought that tumor cells escape immune attack by expressing RCAS1, which induces apoptosis in receptor-positive immune cells. The current study was designed to elucidate the histogenesis of these tumors by using various immunohistochemical markers, and identify parameters that will help to establish the criteria of malignancy in the GIMT. We also discuss the clinicopathological significance of RCAS1 expression in the diagnosis and prognosis of GIMTs. A total of 70 cases of GIMTs were reviewed. Immunohistochemistry was performed between 1990 and 2000, with the avidin-biotin-peroxidase complex method on 3 microm-thick sections of formalin-fixed paraffin-embedded specimens of GIMTs. Antibodies to the following antigens were used: KIT (CD117), CD34 alpha-SMA, Desmin, cytokeratin, S-100 protein, p53, and RCAS1. Recurrence-free survival analysis was done with Stat View-J 5.0 statistical packages. Univariate analysis for a recurrence-free prognosis demonstrated that antibody detection of p53 expression (p=0.0333) and expression of RCAS1 (p=0.0008) is correlated with a significantly higher potential of recurrence. On multivariate analysis, tumor size and RCAS1 expression were independently and inversely correlated with recurrence-free survival. The expression of RCAS1 has not previously been reported in GIMT; indeed, our study suggests that the expression of RCAS1 is correlated with recurrence not only in carcinomas, but also in mesenchymal tumors.

The effects of child formula toothpastes on enamel caries using two in vitro pH-cycling models

To compare, using two pH-cycling models, the de/remineralisation effects of children's toothpastes on primary teeth. In vitro single-section and pH-cycling models. Primary teeth were placed in demineralising solution for 96 hours to produce artificial carious lesions 60-100 microm deep. They were cut into 100 microm thick sections and assigned to 6 groups. Sections in Groups A and D were exposed to a non-fluoridated toothpaste, those in Groups B and E to half-pea-sized (0.16g) and those in Groups C and F to pea-sized portions (0.32g) of a 500ppm F toothpaste. pH-cycling Model I (Groups A, B, C), without added fluoride, ran for 7 days, while Model II (Groups D, E, F), with 0.25ppm F, ran for 10 days. Lesions were evaluated using polarised light microscopy and microradiography. Lesions in Groups B and E progressed by 64% and 61%, respectively, while those in Groups C and F progressed by only 19% and 23% respectively. Both 10-day and 7-day pH-cycling models were suitable for studying carious lesion progression in primary teeth (the demineralising and remineralising solutions of the 10-day cycling model contained 0.25ppm F). A pea-sized portion (0.32g) of 500ppm F toothpaste slowed down the demineralisation progression better than a half-pea-sized portion.

Distribution of chlorophyll-bearing organelles in the shoot apex of a range of dicotyledonous plants

Confocal laser scanning microscopy was used to study the distribution of the smallest detectable autofluorescing, chlorophyll-bearing structures in fresh, 40 microm thick longitudinal sections of the shoot apex of four dicotyledonous plants (Arabidopsis thaliana, Nicotiana glauca, Lupinus alba, and Spinacia oleracea). In all species, the smallest chlorophyll-bearing particles were found in the outermost cell layers (L1 and L2) of the shoot apex. Their distribution between these layers differed in each species. The smallest such particles were about 0.5-1.0 microm in maximum dimension, approximating the size of a single granum in the developing leaf. Their size and abundance increased with increasing cell age and distance from the peak of the apex. Immediately beneath the L1 and L2 layers was a zone largely devoid of these particles. Below this nonfluorescing zone, in the region where the derivatives of the meristematic zone differentiate into cells of the central pith region, the size and abundance of the chlorophyll-bearing particles increased progressively with increasing distance from the nonfluorescing zone. The presence of these small autofluorescing particles in the L1 and L2 cell layers of the shoot apex places the development of photosystem II fluorescence at an earlier stage of leaf development than previously observed. The use of confocal laser scanning microscopy to study unfixed sections provides another useful metabolic marker for mapping patterns of differentiation and development in the cells of the shoot apex.

Preliminary analysis of mutations in exon 5 of IkappaBalpha gene in Hodgkin lymphoma

To study the mutations in exon 5 of IkappaBalpha gene and their significance in Hodgkin lymphoma. CD30-positive Reed-Sternberg cells and surrounding reactive lymphocytes were selected by laser microdissection technology from 6 microm-thick histologic sections of 7 Hodgkin lymphoma cells studied. Genomic DNA of these cells was extracted according to Qiagen micro DNAkit guide. Semi-nested polymerase chain reaction (PCR) and direct DNA sequencing were used to analyze exon 5 of IkappaBalpha gene. A mutation in exon 5 was identified in 3 of the 7 Hodgkin lymphoma cases. No mutation found in the surrounding reactive lymphocytes. The mutation site was located at codon 2084 (T-A, NT_026437), resulting in formation of a stop codon. The mutation identified in exon 5 of the IkappaBalpha gene in Hodgkin lymphoma may play a role in the pathogenesis of Hodgkin lymphoma.

Correlation between TNM classification and malignancy histological feature of oral squamous cell carcinoma

SummaryHistological staging of deep invasive margin of oral squamous cell carcinoma has a significant influence on survival of patients since the tumor cells are more poorly differentiated in this area and have high prognostic value.Aimthe purpose of the present study is to correlate TNM clinical classification with histopathologic characteristics (degree of keratinization, nuclear pleomorphism, invasion pattern and lymphoplasmocytic infiltrate) and histologic malignancy scores in 38 cases of oral epidermoid carcinoma in the lesion's deepest areas.Study FormRetrospective clinical study.Material and MethodThis is a retrospective study based on histological review of 38 cases of oral squamous cell carcinoma selected from the medical files of Hospital Dr. Luis Antonio, Natal - Rio Grande do Norte, Brazil. TNM clinical classification data were obtained from the analysis of the medical records. Two pathologists performed histological malignancy staging on routine 3 µm-thick sections of invasive tumor areas stained with hematoxylin and eosin. For statistical analysis, parametric (ANOVA) and non-parametric tests (Tukey; Pearson; Chi2) were employed.ResultsWe found significant correlation between TNM clinical staging and malignancy mean score (p = 0.001) and histopathologic parameters, such as nuclear pleomorphism (p = 0.016) and degree of keratinization (p = 0.025). Furthermore, there were also statistically significant correlations between lymphocytic infiltration (p = 0.016) and nuclear pleomorphism (p = 0.004) with TNM classification when grouped in two series: TNM I/II and III/IV.ConclusionTNM classification, as well as malignancy mean score, had statistically significant correlation with degree of keratinization, nuclear pleomorphism and lymphocytic infiltration. These highly significant results indicated that histologically invasive areas may be primarily responsible for the clinical behavior of the tumor, and this may be important for the therapy of choice for oral squamous cell carcinoma.

Multiple metastases to the small bowel from large cell bronchial carcinomas

Metastases from lung cancer to gastrointestinal tract are not rare at postmortem studies but the development of clinically significant symptoms from the gastrointestinal metastases is very unusual. Formalin-fixed, paraffin-embedded tissues were cut into 5 microm thick sections and routinely stained with hematoxylin and eosin. Some slides were also stained with Alcian-PAS. Antibodies used were primary antibodies to pancytokeratin, cytokeratin 7, cytokeratin 20, epithelial membrane antigen, vimentin, smooth muscle actin and CD-117. We observed three patients who presented with multiple metastases from large cell bronchial carcinoma to small intestine. Two of them had abdominal symptoms (sudden onset of abdominal pain, constipation and vomiting) and in one case the tumor was incidentally found during autopsy. Microscopically, all tumors showed a same histological pattern and consisted almost exclusively of strands and sheets of poorly cohesive, polymorphic giant cells with scanty, delicate stromas. Few smaller polygonal anaplastic cells dispersed between polymorphic giant cells, were also observed. Immunohistochemistry showed positive staining of the tumor cells with cytokeratin and vimentin. Microscopically and immunohistochemically all metastases had a similar pattern to primary anaplastic carcinoma of the small intestine. In patients with small intestine tumors showing anaplastic features, especially with multiple tumors, metastases from large cell bronchial carcinoma should be first excluded, because it seems that they are more common than expected.