Nuclear Vacuoles Research Articles

GINGIVAL ANGIOLEIOMYOMA IN CHILD

Male patient, 10 years old, was referred to evaluate symptomatic lesion with duration of 2 months. There were no systemic diseases. Extraoral and intraoral examinations revealed nodular soft tissue mass located in the upper gingival, between 63 and 24, covered for normal oral mucosa with ulceration area. Periapical and panoramic radiography showed smaller radiopacity of the alveolar bone around the root of 63. Diagnostic hypothesis were pyogenic granuloma and giant cells peripheral lesion. Excisional biopsies and root planning of associated teeth were performed. Histologic analysis revealed spindle-shaped cells with eosinophilic cytoplasm and elongated blunt-ended cigar-shaped nuclei to large dilated vascular channels. Masson's trichrome stain and immunohistochemistry were performed nuclear vacuole to differentiate angioleiomyoma and myofibroma. Diagnosis was angioleiomyoma. The patient developed no recurrence in the thirty months after surgical treatment. In this case, Masson's trichrome stain demonstrated to be an important method to differentiate angioleiomyoma and myofibroma. (CNPq #309322/2015-4; FAPEMIG).

Systemic effects of H2S inhalation at human equivalent dose of pathologic halitosis on rats

Objectives: Halitosis is composed by hundreds of toxic gases. It is still not clear whether halitosis gases self-inhaled by halitosis patients cause side effects. The aim of the study was to investigate the effect of H2S inhalation at a low concentration (human equivalent dose of pathologic halitosis) on rats.Materials and methods: The threshold level of pathologic halitosis perceived by humans at 250 ppb of H2S was converted to rat equivalent concentration (4.15 ppm). In the experimental group, 8 rats were exposed to H2S via continuous inhalation but not the control rats. After 50 days, blood parameters were measured and tissue samples were obtained from the brain, kidney and liver and examined histopathologically to determine any systemic effect.Results: While aspartate transaminase, creatine kinase-MB and lactate dehydrogenase levels were found to be significantly elevated, carbondioxide and alkaline phosphatase were decreased in experimental rats. Other blood parameters were not changed significantly. Experimental rats lost weight and became anxious.Histopathological examination showed mononuclear inflammatory cell invasion in the portal areas, nuclear glycogen vacuoles in the parenchymal area, single-cell necrosis in a few foci, clear expansion in the central hepatic vein and sinusoids, hyperplasia in Kupffer cells and potential fibrous tissue expansion in the portal areas in the experimental rats. However, no considerable histologic damage was observed in the brain and kidney specimens.Conclusions: It can be concluded that H2S inhalation equivalent to pathologic halitosis producing level in humans may lead to systemic effects, particularly heart or liver damage in rats.

Cytomorphological changes induced by intraperitoneal chemotherapy present important diagnostic pitfalls in peritoneal fluid cytology

Intraperitoneal chemotherapy (IP ChT) is an emerging modality in the treatment of advanced gastric adenocarcinoma with peritoneal disease. Cytological evaluation of peritoneal fluid specimens from patients undergoing IP ChT is important in clinical management. However, direct intraperitoneal exposure to chemotherapeutic agents induces cytomorphological changes in benign constituents of peritoneal fluid, presenting particular challenges to accurate cytological interpretation. These morphological changes have not been well characterised in the literature. We systematically reviewed the cytomorphological features in immunocytochemically-confirmed positive and negative IP ChT peritoneal fluid samples to elucidate the degree of morphological overlap between malignant and reactive cells. We reviewed 39 peritoneal fluid samples of patients treated with IP ChT, and scored specific cytomorphological parameters of both benign and malignant cells with the aid of relevant immunocytochemical interrogation. The present findings show a significant degree of morphological overlap between reactive and malignant cells. Abnormal, "exploding" mitotic figures, nuclear membrane irregularities, multi-nucleation and cytoplasmic vacuolation were commonly observed in negative fluid specimens. The most helpful feature that favoured malignant cells was the increased nuclear-to-cytoplasmic ratio. A background inflammatory milieu of eosinophils and/or neutrophils was seen in 45-58% of post IP ChT peritoneal fluid specimens. The presence of pseudoparakeratotic cells, a novel observation in post IP ChT fluid specimens is also described. The extent of reactive cytomorphological anomalies arising from treatment with IP ChT poses unique diagnostic challenges and may prompt a malignant or 'atypical' diagnosis in benign reactive samples.

Cytopathologic evaluation of patients submitted to radiotherapy for uterine cervix cancer.

Cervical cancer is an important public health problem. Pap smear is the leading strategy of screening programs for cervical cancer worldwide. However, delayed diagnosis leads to more aggressive and less effective treatments. Patients with uterine cervix malignancies who are referred for radiotherapy have advanced-stage disease, which results in high rates of locoregional recurrence. The use of radiotherapy as a treatment for cervical cancer causes morphological changes in neoplastic and non-neoplastic epithelial cells, as well as in stromal cells, which make it difficult to diagnose the residual lesion, resulting in a dilemma in cytopathological routine. Based on the difficulties of cytopathologic evaluation for the follow-up of patients treated with radiotherapy for cervical cancer, our objective was to describe the actinic cytopathic effects. Our paper was based on a structured review including the period from June 2015 to April 2016, aiming at an exploratory-descriptive study. Bibliographic investigations were carried out through selection and analysis of articles, list of authors and keywords, selection of new articles focused on the analysis of bibliographic references to previously selected documents, as well as textbooks of recognized merit. The most incident actinic cytopathological alterations as described in the literature are: cellular gigantism, nuclear and cytoplasmic vacuolization, dyskeratosis, bi- and multinucleated (B/M) cells, macro and multiple nucleoli, anisokaryosis, anisonucleolosis and nuclear pyknosis. To date, a protocol has not been established that can precisely differentiate the morphological characteristics between benign cells with actinic effects from recurrent malignant cells on post-radiotherapy smears.

An investigation of the potential effect of sperm nuclear vacuoles in human spermatozoa on DNA fragmentation using a neutral and alkaline Comet assay

Presence of vacuoles and degree of sperm DNA damage are considered to be the basic factors used for the assessment of sperm fertilization capacity. We aimed to investigate the link between these two parameters. According to our knowledge, this is the first study where the Comet assay was used to assess the degree of DNA fragmentation of sperm categorized by Motile Sperm Organelle Morphology Examination (MSOME) Grades. Semen samples from 10 patients were assessed. Spermatozoa were graded into four MSOME groups according to the Vanderzwalmen's criteria. A total of 3930 motile spermatozoa were selected one-by-one using an inverted microscope and transferred onto two different slides. The degree of DNA fragmentation was analyzed by alkaline and neutral Comet assay. Results of the neutral Comet assay showed that Grade I spermatozoa (absence of vacuoles) presented significantly lower dsDNA fragmentation level (mean: 3.13±1.17%) than Grade II (maximum of two small vacuoles; mean: 10.34±2.65%), Grade III (more than two small vacuoles or at least one large vacuole; mean: 23.88±8.37%), and Grade IV (large vacuoles associated with abnormal head shapes or other abnormalities; mean: 36.94±7.78%; p<0.05). Results of the alkaline Comet assay showed that Grade I spermatozoa had significantly lower DNA (ssDNA+dsDNA) fragmentation level (mean: 8.33±3.62%) than Grade III (mean: 25.64±9.15%) and Grade IV (mean: 40.10±9.10%, p<0.05), but not significantly lower than Grade II (mean: 12.73±5.06%; p>0.05). Probably, the vacuoles may be responsible for double strand DNA breaks rather than single strand DNA breaks (only 2.39% spermatozoa in MSOME Grade II, 1.76% in III, and 3.16% in IV has single strand breaks). The results demonstrate that lower MSOME grading correlates with lower sperm DNA fragmentation. Therefore, the observation of sperm nuclear vacuoles using real-time optical microscopy without precise DNA fragmentation examination is not sufficient for optimal sperm selection for intracytoplasmic sperm injection.

High prevalence of MiTF staining in undifferentiated pleomorphic sarcoma: caution in the use of melanocytic markers in sarcoma.

The diagnosis of undifferentiated pleomorphic sarcoma (UPS) may be challenging, as other lesions with undifferentiated spindle cell morphology must be excluded, including melanoma. Microphthalmia-associated transcription factor (MiTF) stains naevi and epithelioid melanomas, as well as some mesenchymal neoplasms. The aim of this study was to evaluate the prevalence of MiTF and melanocytic markers in UPS and a subset of atypical fibroxanthoma (AFX). MiTF, SOX10, Melan-A, HMB45 and S100 immunostaining was performed on resection specimens from 19 UPSs and five AFXs. Next-generation sequencing of 50 genes was performed in UPSs to exclude dedifferentiated melanoma. In 17 of 19 UPSs (89%), tumour cells showed nuclear positivity for MiTF that was not eliminated by casein block. Three showed focal nuclear staining for HMB45, which was eliminated by casein block. One showed focal nuclear vacuole staining for S100 with red but not brown chromogen. None expressed SOX10 or Melan-A. Mutational analysis of 15 UPSs with adequate DNA showed no mutations within hotspot regions of BRAF, KIT, or NRAS. Four of five AFXs (80%) stained with MiTF; other markers were negative. There is a high prevalence of nuclear MiTF expression in UPSs (89%) and AFXs (80%). Rare UPSs showed non-specific nuclear HMB45 or S100 staining. These findings argue against using MiTF in isolation to differentiate between UPS or AFX and melanoma, and caution in interpreting focal staining for a single additional melanocytic marker. Casein block may eliminate non-specific staining. MiTF should be used to support a diagnosis of melanoma only if multiple melanocytic markers are positive.

Cytomorphometric analysis and morphological assessment of oral exfoliated cells in type 2 diabetes mellitus and healthy individuals: A comparative study.

Context:Oral exfoliative cytology is a simple, nonaggressive technique that is well accepted by patients. Therefore, it is an attractive option, which aids in the diagnosis and observation of epithelial atypias associated with oral mucosal diseases.Aims:The aim of this study was to evaluate and compare the quantitative and qualitative alterations in exfoliative smears from type 2 diabetics and healthy individuals.Patients and Methods:The study includes 30 type 2 diabetics and 30 healthy persons of both sexes. PAP and hematoxylin and eosin (H and E) stained smears were prepared from buccal mucosa (BM), tongue (T), floor of the mouth (FOM), and palate (P). Under a light microscope, 50 clearly defined unfolded epithelial cells were quantitatively evaluated for cellular area (CA), nuclear area (NA), and cellular-to-nuclear area ratio (CA:NA) and assessed for morphological features.Statistical Analysis:Collected data was manually entered into the Statistical Package for the Social Sciences version 13.5 for analysis. Student's t-test was used at 95% confidence interval.Results:Quantitative assessment of the overall mean CA was less, mean NA was more, and mean CA:NA was less in diabetics than that in healthy persons at all the four sites. Diabetic oral cells showed qualiative cytoplasmic and nuclear alterations: cytoplasmic vacuoles, karyorrhexis, karyolysis, pyknosis, peri-nuclear halo, binucleation, nuclear vacuoles, inflammation, and microbial colonies.Conclusion:Oral cytology from type 2 diabetics is associated with detectable cytomorphological changes with alteration in size of the cell and nucleus, which is site specific, indicating epithelial cell degeneration in cytoplasm and nucleus.

Clinical and Immunohistochemical Features of Sebaceous Carcinoma: Focusing on the p53 Tumor Suppressor

Purpose: Considerable focus has recently revealed the importance of p53 in sebaceous carcinoma (SC). This study investigates the utility of p53 as an immunohistochemistry (IHC) marker for differentiating benign and malignant sebaceous tissues and reports the clinical characteristics of patients diagnosed with SC at Mount Sinai Hospital (MSH). Methods: A retrospective chart analysis of 102 patients with SC was performed in addition to a histopathological examination of benign and malignant sebaceous tissues. Immunohistochemistry was performed in 30 cases of SC, 4 cases of sebaceous hyperplasia, 4 cases of sebaceous epitheliomas, and 3 cases of sebaceous adenoma. A separate outpatient cohort was used to compare the clinical and histologic features of eyelid lesions. This included 31 available cases with histologic descriptions of SC. The clinical features of 20 SC cases, 25 BCC cases, and 21 chalazia cases were available and compared from this outpatient cohort. Data collected included age, gender, lesion anatomical location, histopathologic descriptions, and percentage of p53 positive sebocytes. Results: Of the 102 SC cases at MSH, the mean age was 69 years, males composed 58%, and 59% were periorbital or orbital. Positive p53 staining without atypia was reproducibly observed at cell membranes in all cases of sebaceous hyperplasia. Using a standardized p53 grading system used at MSH, the average p53 staining grade for SC and epithelioma was calculated to be 2.3 ± 0.7 and 1 ± 0.7, respectively (p=0.002). An outpatient cohort revealed that 65% of SC was found on the lower eyelid and 84% of BCC was on the upper eyelid. The most common histologic features seen in SC were nuclear atypia (61%) and cytoplasmic vacuoles (55%). Conclusions: This study provides the first comprehensive staining profile of p53 in benign and malignant sebaceous tissues. We also report for the first time that membrane p53 staining was helpful and appeared to be highly specific for identifying sebaceous hyperplasia. p53 staining was also helpful for identifying SC and intraepithelial spreading tumor cells. The clinical and histologic characteristics of specific eyelid lesions in an outpatient setting are also shared. Lastly, this work incorporates the latest research to synthesize IHC recommendations for improving SC detection.

Mechanism of grape seeds extract protection against paracetamol renal cortical damage in male Albino rats.

The aim of the study was to assess the possible protective role of grape seeds extract (GSE) in ameliorating the toxic effects of paracetamol overdose on the rat renal cortical tissue. Paracetamol is one of the widely used non-steroidal anti-inflammatory drugs (NSAIDs). Unfortunately, it was reported as the most common cause of toxic ingestion in the world. Grape seeds extract (GSE) is known to have a strong antioxidant and anti-inflammatory properties. The rats were divided into 4 groups; control group, GSE group, paracetamol group and GSE with paracetamol group. Kidney specimens were processed for biochemical, histological and immunohisto-chemical studies. The study showed marked biological changes in the form of significant increase in serum urea and creatinine levels with significant decrease in renal superoxide dismutase with paracetamol group. Furthermore, Proximal (PCT) and distal convoluted tubules showed marked degeneration, dense nuclear staining, cytoplasmic vacuolization, and partial loss of the brush borders. Most tubules were dilated, irregular and were filled with hyaline casts. PCT and DCT showed less PAS reaction and more COX-2 and caspase expression if compared with the control and the GSE groups. Concomitant administration of grape seeds extract with paracetamol revealed a noticeable amelioration of these biochemical and histological changes. Proximal and distal convoluted tubules showed less PAS reaction and more COX2 and caspase expression if compared with the control and the GSE. Concomitant administration of GSE with paracetamol revealed a noticeable amelioration of these biochemical and histological changes. Grape seeds extract provided biochemical and histo-pathological improvement in paracetamol induced renal cortical toxicity. These findings revealed that this improvement was associated with a decrease in oxidative damage and apoptosis (Tab. 1, Fig. 7, Ref. 55).

Predicting the clinical outcome of ICSI by sperm head vacuole examination

ABSTRACTTo assess whether high magnification sperm head vacuole examination (SHVE) and/or standard sperm morphology assessment can predict ICSI outcomes in terms of fertilization, embryo quality, and delivery rates, a prospective observational bicentric study was conducted in two publicly funded assisted reproductive technology (ART) units in France between January and July of 2012. A total of 111 ICSI cycles for exclusively male infertility factors were included. A Spearman’s correlation test was performed to validate SHVE reproducibility between the ART units. The normal morphology rate and SHVE performed on selected spermatozoa were respectively determined according to David’s and Vanderzwalmen’s classifications used for motile sperm organelle morphology examination (MSOME) on the day of the ICSI. Receiver Operating Characteristic (ROC) curve analysis was performed to determine thresholds associated with the occurrence of a delivery. There was an excellent correlation between the two operators (r=0.98), thus validating the study’s SHVE data. Percentages of normal morphology grade spermatozoa using the standard classification and first-best morphology grade spermatozoa determined by SHVE were not significantly associated with (i) delivery (p=0.58; 0.90 /area under curve (AUC) =0.532; 0.507), (ii) fertilization (p=0.88; 0.90), (iii) top-quality embryos (p=0.27; 0.98), and (iv) good quality embryo rates (p=0.73; 0.98), respectively. In conclusion, high magnification SHVE and standard sperm morphology assessment cannot predict clinical or biological ICSI outcomes.Abbreviations: ART: assisted reproductive technology; HBV: hepatitis B virus; HCV: hepatitis C virus; HIV: human immunodeficiency virus; ICSI: intra-cytoplasmic sperm injection; IVF: in vitro fertilization; LNVs: large nuclear vacuoles; MSOME: motile sperm organelle morphology examination; SHVE: sperm head vacuole examination; WHO: World Health Organization

Npa3/ScGpn1 carboxy-terminal tail is dispensable for cell viability and RNA polymerase II nuclear targeting but critical for microtubule stability and function

Genetic deletion of the essential GTPase Gpn1 or replacement of the endogenous gene by partial loss of function mutants in yeast is associated with multiple cellular phenotypes, including in all cases a marked cytoplasmic retention of RNA polymerase II (RNAPII). Global inhibition of RNAPII-mediated transcription due to malfunction of Gpn1 precludes the identification and study of other cellular function(s) for this GTPase. In contrast to the single Gpn protein present in Archaea, eukaryotic Gpn1 possesses an extension of approximately 100 amino acids at the C-terminal end of the GTPase domain. To determine the importance of this C-terminal extension in Saccharomyces cerevisiae Gpn1, we generated yeast strains expressing either C-terminal truncated (gpn1ΔC) or full-length ScGpn1. We found that ScGpn1ΔC was retained in the cell nucleus, an event physiologically relevant as gpn1ΔC cells contained a higher nuclear fraction of the RNAPII CTD phosphatase Rtr1. gpn1ΔC cells displayed an increased size, a delay in mitosis exit, and an increased sensitivity to the microtubule polymerization inhibitor benomyl at the cell proliferation level and two cellular events that depend on microtubule function: RNAPII nuclear targeting and vacuole integrity. These phenotypes were not caused by inhibition of RNAPII, as in gpn1ΔC cells RNAPII nuclear targeting and transcriptional activity were unaffected. These data, combined with our description here of a genetic interaction between GPN1 and BIK1, a microtubule plus-end tracking protein with a mitotic function, strongly suggest that the ScGpn1 C-terminal tail plays a critical role in microtubule dynamics and mitotic progression in an RNAPII-independent manner.

Cytotoxic and apoptotic activities of extract of Amaranthus spinosus L. in Allium cepa and human erythrocytes.

The present study examined the apoptosis inducing effects of Amaranthus spinosus L. aqueous extract in Allium cepa root meristematic cells and human erythrocytes. Cytogenetic assay revealed many apoptosis inducing cytogenetic aberrations viz., cytoplasmic breakage, cytoplasmic disintegration, cytoplasmic shrinkage, receding of cytoplasm, cytoplasmic vacuolation, enucleated cell, ghost cell, nuclear vacuolation, nuclear fragmentation and nuclear disintegration. A remarkable modification of red blood cell surface morphology was observed in the result of RBC assay. The treated RBCs show membrane blebbing and shrinkage, features typical for apoptosis in nucleated cells. Significant induction of cell death was observed in treated Allium root tip cells after Evans blue staining, disclosing the membrane damage potential of the plant extract. TTC assay results in reduced mitochondrial/metabolic activity in Allium root tip cells after treatment, designating the adverse effect of plant extract on mitochondrial respiratory chain. These results confirm the apoptosis inducing potential of A. spinosus extract. Confirming the present results by further in vitro studies, it can be effectively targeted against cell proliferation during cancer treatment by inducing apoptosis. Thus from the present investigation it can be concluded that the aqueous extract of A. spinosus exhibited apoptosis induction and cytotoxic activities.

English

Control of leishmaniasis remains a challenge due the high toxicity of the chemotherapeutic drugs presently available. The ongoing search for better leishmanicidal compounds has brought herbal drugs into the limelight as safe and effective substitutes to conventional therapies which have various drawbacks. The current study was designed to evaluate the antileishmanial effect of Quassia amara against Leishmania amazonensis and Leishmania infantum. Different fractions (Hexane (Q1), dichloromethane (Q2), ethyl acetate (Q3) and butanol (Q4)) obtained from the liquid-liquid partition of the crude Q. amara methanol extract were tested against Leishmania promastigote forms. After 120 h of treatment with Q. amara fractions (1 to 500 &micro;g/ml), the minimal inhibitory concentration (MIC) was determined using resazurin. The most active fraction (Q2) was analyzed using thin layer chromatography (TLC) and high performance liquid chromatography-diode array detector (HPLC-DAD) techniques. The effects of Q2 on parasite ultrastructure were investigated by transmission electron microscopy. In addition, Leishmania-infected macrophages were treated with Q2 in order to evaluate the anti-amastigote effect. Among the fractions tested, Q2 showed the highest activity against L. amazonensis and L. infantum promastigotes MIC values of 62.5 and 31.25 &micro;g/ml, respectively. There were ultrastructural alterations, such as nuclear chromatin condensation, intense vacuolization and autophagic and myelin-like figures in parasites treated with Q2 (62.5 &micro;g/ml). Macrophages previously infected with L. amazonensis and L. infantum promastigotes showed a drastic reduction in the number of parasites recovered in the supernatant after Q2 treatment at MIC values. The TLC and HPLC fingerprints of Q2 showed that alkaloids were the main chemical constituents in this fraction. The results presented herein showed that the alkaloid-rich fraction Q2 is a promising source of antileishmanial agents. Further investigation will be necessary in order to isolate and test the substance(s) responsible for the bioactivity. Key words: Antileishmanial activity, Quassia genus, alkaloids, macrophage infection, nitric oxide.

Retinal toxicity by intravitreal liquid perfluorocarbon

PurposeTo report retinal changes in a population of patients who had undergone vitreoretinal surgery using liquid perfluorocarbon (LPFC).MethodsSingle centre, retrospective study. The clinical records and images of patients who had undergone vitreoretinal surgery during the 36 months prior to the sanitary alert issued by the Spanish Health Authorities on ALA OCTA LPFC were revised.ResultsFour different brands of LPFC had been used during the 36 months period, including ALA OCTA. 128 patients were identified who had undergone vitreoretinal surgery with LPFC of whom 44 had been treated with ALA OCTA. None of the eyes fulfilled the diagnostic criteria of the sanitary alert issued by the Spanish Health Authorities (no light perception + retinal necrosis + optic disk atrophy). No sign of retinal necrosis or severe ocular inflammation was identified in any of the eyes treated with any of the brands during postoperative follow‐up or at the final visit. Optic disk atrophy was identified in 6 eyes, and limited venous sheathing was observed in one eye treated with ALA OCTA vs. no cases in eyes treated with other brands. SDOCT revealed retinal ganglion cell layer vacuolization in eight eyes and inner nuclear layer vacuolization in five eyes treated with ALA OCTA.ConclusionsA range of retinal lesions were identified in patients operated on with the aid of LPFC. These changes and lower final visual acuities were more frequently observed but were not exclusive among eyes treated by ALAOCTA LPFC. LPFC toxicity must be carefully evaluated in a case‐by‐case basis in order to elucidate the role of LPFC and other factors involved

Yeast Vps13 promotes mitochondrial function and is localized at membrane contact sites.

The Vps13 protein family is highly conserved in eukaryotic cells. Mutations in human VPS13 genes result in a variety of diseases, such as chorea acanthocytosis (ChAc), but the cellular functions of Vps13 proteins are not well defined. In yeast, there is a single VPS13 orthologue, which is required for at least two different processes: protein sorting to the vacuole and sporulation. This study demonstrates that VPS13 is also important for mitochondrial integrity. In addition to preventing transfer of DNA from the mitochondrion to the nucleus, VPS13 suppresses mitophagy and functions in parallel with the endoplasmic reticulum-mitochondrion encounter structure (ERMES). In different growth conditions, Vps13 localizes to endosome-mitochondrion contacts and to the nuclear-vacuole junctions, indicating that Vps13 may function at membrane contact sites. The ability of VPS13 to compensate for the absence of ERMES correlates with its intracellular distribution. We propose that Vps13 is present at multiple membrane contact sites and that separation-of-function mutants are due to loss of Vps13 at specific junctions. Introduction of VPS13A mutations identified in ChAc patients at cognate sites in yeast VPS13 are specifically defective in compensating for the lack of ERMES, suggesting that mitochondrial dysfunction might be the basis for ChAc.

Sperm nuclear vacuoles impair the reproductive success in a human in vitro fertilization program (IVF)

Sperm nuclear vacuoles impair the reproductive success in a human in vitro fertilization program (IVF)

Adenoid ameloblastoma with dentinoid and cellular atypia: a rare case report

Adenomatoid odontogenic tumor (AOT) is always a benign tumor with rare incidence of recurrence while ameloblastoma is the commonest gnathic tumor, which is always aggressive. Although co-occurrence of these lesions has been reported, this paper describes a homogenous combination of &lt;em&gt;atypical&lt;/em&gt; AOT and ameloblastomatous proliferation with some malignant microscopic features. To date, A dozen cases or slightly more of this uncommon composite odontogenic tumor have been, quite correctly, reported in the literature under the designation of &lt;em&gt;adenoid ameloblastoma&lt;/em&gt;. Of these, neither cellular atypia nor pleomorphism has been revealed. This extremely rare ameloblastomatous variant can pose a significant diagnostic challenge. Moreover, there were some findings of severe nuclear vacuolization, mitotic figures, cellular pleomorphism and nuclear hyperchromatism and chromatin peripheralization. However, the scattered occurrence of these was not sufficient for claiming a malignancy. To confirm, two immunohistochemical markers - Calretinin and p53 - were recruited. Rendering itself to be suspicious, a rapt attention should be paid toward well interrogating this lesion histologically and immunohistochemically.

Abstract WMP111: Stroke and Alzheimer’s Disease: Shared Mechanisms of Vascular Disruption

Recent evidence suggests that Alzheimer’s disease is a vascular disorder that can be accelerated by ischemic stroke. We propose that the underlying pathophysiology accelerating degenerative changes is due to blood brain barrier disruption following ischemia. A transient middle cerebral artery occlusion suture model was used in triple-transgenic AD mice and C57/Bl6 wild type controls. A separate triple-transgenic AD mice group was used for sham control. The middle cerebral artery was occluded for 1 hour followed by reperfusion. Mice were evaluated for sickness behavior and received a modified neurologic score. 5 hours after reperfusion, blood brain barrier tracers (Texas red, IcG, and AIB1) were injected i.v. into the femoral vein and allowed to circulate for 10 minutes. H&amp;E, Cresyl Violet, and Thioflavin were used to stain sections of the brain for areas of degeneration. Images were obtained by quantitative fluorescent and bright-field microscopy. One-way ANOVA with Tukey’s post-hoc comparison was used to compare between groups. P&lt;0.05 was considered statistically significant. A statistically significant difference between groups was seen for blood brain barrier permeability following ischemia and reperfusion. Triple-transgenic AD mice had a 2-fold increase in Texas red and a 4-fold increase in AIB1 at 5 hours after reperfusion. No significant differences were seen in the control groups. H&amp;E staining revealed nuclear fragmentation, chromatolysis, and vacuolization in the ipsilateral cortex of the triple-transgenic AD mice exposed to stroke. Limited areas of necrosis were seen in the C57/Bl6 controls. A significant increase in thioflavin staining was seen post-stroke in the triple-transgenic AD mice. The amyloid distribution was perivascular and significantly increased compared to the triple-transgenic AD sham mice. Triple-transgenic AD mice had a worse sickness score (avg. 13) compared to C57/Bl6 controls (avg. 17). Stroke and Alzheimer’s disease share similar vascular mechanisms related to blood brain barrier disruption. We show that blood brain barrier disruption post-stroke accelerates degenerative changes. Targeting blood brain barrier disruption as a treatment approach may prove promising and warrants continued investigation.

Hepatic Parameters of Marine Fish Rachycentron canadum (Linnaeus, 1766) Exposed to Sublethal Concentrations of Water-Soluble Fraction of Petroleum

Hepatic Parameters of Marine Fish Rachycentron canadum (Linnaeus, 1766) Exposed to Sublethal Concentrations of Water-Soluble Fraction of Petroleum The aim of this paper was to evaluate the liver histological changes, hepatosomatic index and condition factor, as well as the activity of the enzymes alanine aminotransferase and aspartate aminotransferase after sublethal exposure 0.3 ppm of water-soluble fraction (WSF) of petroleum the period from 7 and 14 days. Observed an increase of hepatosomatic index after 7 and 14 days and reduced condition factor at 14 days of exposure, increased AST activity in serum at 14 days of exposure and liver histological alterations in 7 and 14 days, such as hypertrophy of nucleus of the hepatocytes, increase of density of the hepatic blood vessels, disarrangement of hepatic cords, cell atypia contour, intense cytoplasmic and nuclear vacuolization and hyperemia. We concluded that the evaluation of hepatosomatic index, histopathological alterations and activity of enzymes aspartate aminotransferase (AST) are sensitive to exposure to WSF, being good biomarkers. It is concluded, also, that the sub-lethal exposure to 0.3ppm WSF change of hepatic parameters of the fish species Rachycentron canadum, affecting the functioning of the biotransformation system. Knowledge of the changes and their adaptations will serve as the basis for the conservation of the species and more effective environmental monitoring

Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms.

Density gradient centrifugation (DGC) and swim-up techniques have been reported for semen preparation in assisted reproductive techniques in humans. We investigated whether semen preparation using a combination of DGC and swim-up techniques could effectively decrease morphologically abnormal human sperms at the ultrastructural level. Semen samples were obtained from 16 infertile males and fractionated by swim-up following DGC. Ultrastructural abnormalities of sperms obtained from original semen, lower layer of swim-up following DGC, and upper layer of swim-up following DGC were analyzed by transmission electron microscopy. The correlation among ultrastructural head abnormality in sperms from the upper layer of swim-up, fertilization in in vitro fertilization, and pregnancy after embryo transfer was also investigated. Furthermore, sperms with DNA fragmentation in the samples processed via a combination of DGC and swim-up was assessed in a sperm chromatin structure assay. Ultrastructural abnormalities in sperm heads and tails in the upper layer after swim-up following DGC was the lowest among the three groups. Sperms with nuclear vacuoles were the most difficult to eliminate using a combination of DGC and swim-up in all types of head abnormalities. A negative correlation was confirmed between the fertilization rates of intracytoplasmic sperm injection and head abnormality of sperms obtained from the upper layer of the swim-up following DGC. Sperms with DNA fragmentation were effectively decreased using the combination of two techniques. In conclusion, the combination of DGC and swim-up effectively decreased the number of sperms with ultrastructural abnormalities both in the head and in the tail. However, sperms with ultrastructural abnormalities that cannot be completely decreased using a combination of DGC and swim-up may impair fertilization in some cases of intracytoplasmic sperm injection.