Histone deacetylase inhibitor treatment restores memory-related gene expression and learning ability in neonicotinoid-treated Apis mellifera.

Apis mellifera plays crucial roles in maintaining the balance of global ecosystems and stability of agricultural systems by helping pollination of flowering plants, including many crops. In recent years, this balance has been disrupted greatly by some pesticides, which results in great losses of honeybees worldwide. Previous studies have found that pesticide-caused memory loss might be one of the major reasons for colony loss. Histone deacetylase inhibitors (HDACis) are chemical compounds that inhibit the activity of histone deacetylases and are known to cause hyperacetylation of histone cores and influence gene expression. In our study, the HDACi sodium butyrate was applied to honeybees as a dietary supplement. The effect of sodium butyrate on the expression profiles of memory-related genes was analysed by quantitative reverse transcription PCR. The results revealed that this HDACi had up-regulation effects on most of the memory-related genes in bees, even in bees treated with imidacloprid. In addition, using the proboscis extension reflex to evaluate olfactory learning in bees, we found that this HDACi boosted the memory formation of bees after impairment owing to imidacloprid exposure. This study investigated the association between gene expression and memory formation from an epigenetic perspective. Additionally, we further demonstrate the possibility of enhancing bee learning using HDACis and provide initial data for future research.

Chaperones and intimate physical examinations: what do male and female patients want?

INTRODUCTION: Many studies of patients' perception of a medical chaperone have focused on female patients; that of male patients are less well studied. Moreover, previous studies were largely based on patient populations in English-speaking countries. Therefore, this study was conducted to investigate the perception and attitude of male and female Chinese patients to the presence of a chaperone during an intimate physical examination. METHODS: A cross-sectional guided questionnaire survey was conducted on a convenient sample of 150 patients at a public teaching hospital in Hong Kong. RESULTS: Over 90% of the participants considered the presence of a chaperone appropriate during intimate physical examination, and 84% felt that doctors, irrespective of gender, should always request the presence of a chaperone. The most commonly cited reasons included the availability of an objective account should any legal issue arise, protection against sexual harassment, and to provide psychological support. This contrasted with the experience of those who had previously undergone an intimate physical examination of whom only 72.6% of women and 35.7% of men had reportedly been chaperoned. Among female participants, 75.0% preferred to be chaperoned during an intimate physical examination by a male doctor, and 28.6% would still prefer to be chaperoned when being examined by a female doctor. Among male participants, over 50% indicated no specific preference but a substantial minority reported a preference for chaperoned examination (21.2% for male doctor and 25.8% for female doctor). CONCLUSIONS: Patients in Hong Kong have a high degree of acceptance and expectations about the role of a medical chaperone. Both female and male patients prefer such practice regardless of physician gender. Doctors are strongly encouraged to discuss the issue openly with their patients before they conduct any intimate physical examination.

Factors influencing different types of malocclusion and arch form-A review.

AIM: This review intends to highlight malocclusion as a multifactorial issue and review the different factors that influence different types of malocclusion and arch form. METHODS: An online article search was performed on the factors influencing malocclusion and arch form from January 1990 through April 2020. The search was performed within the Google, Rutgers library, PubMed, MEDLINE databases via OVID using the keywords mentioned in the PubMed and MeSH headings for the English language published articles January 1990 through April 2020, which evaluated different factors that influence malocclusion and arch form. RESULTS: Of the 300 articles found in initial search results, 31 articles met the inclusion criteria set for this review. These 31 studies were directly related to the factors that impact malocclusion and different arch forms. CONCLUSION: Genetic inheritance, genetic mutations, age, gender, ethnicity, dental anomalies like macrodontia, congenital diseases, muscular diseases, hormone imbalance, and human behaviour are all factors that influence malocclusion and arch forms. The elements within the individual's control like behaviours can aid in preventing malocclusion. However, it seems as if the underlying reason for most of these factors indicates that malocclusion is a by-product of genetics and pathology.

Automated 3D-printed unibody immunoarray for chemiluminescence detection of cancer biomarker proteins.

A low cost three-dimensional (3D) printed clear plastic microfluidic device was fabricated for fast, low cost automated protein detection. The unibody device features three reagent reservoirs, an efficient 3D network for passive mixing, and an optically transparent detection chamber housing a glass capture antibody array for measuring chemiluminescence output with a CCD camera. Sandwich type assays were built onto the glass arrays using a multi-labeled detection antibody-polyHRP (HRP = horseradish peroxidase). Total assay time was ∼30 min in a complete automated assay employing a programmable syringe pump so that the protocol required minimal operator intervention. The device was used for multiplexed detection of prostate cancer biomarker proteins prostate specific antigen (PSA) and platelet factor 4 (PF-4). Detection limits of 0.5 pg mL-1 were achieved for these proteins in diluted serum with log dynamic ranges of four orders of magnitude. Good accuracy vs. ELISA was validated by analyzing human serum samples. This prototype device holds good promise for further development as a point-of-care cancer diagnostics tool.

Involvement of heregulin-beta2 in the acquisition of the hormone-independent phenotype of breast cancer cells.

The erbB-2 receptor plays an important role in the prognosis of breast cancer. Amplification or overexpression of the erbB-2 proto-oncogene has been detected in 30% of breast cancers and is associated with poor patient prognosis. The significance of erbB-3 and erbB-4 in breast cancer is not yet known. The discovery of the growth factor heregulin (HRG) has allowed us to investigate a number of biological events that are regulated by erbB-2, -3, and -4 signal transduction. To determine the role of HRG in breast cancer tumor progression, we have developed an in vitro/in vivo model. We transfected HRG cDNA into the estrogen receptor (ER)-positive breast cancer cell line, MCF-7, and studied these cells as they progressed from a hormone-dependent to -independent phenotype. The biochemical and biological characteristics presented here demonstrate that overexpression of HRG induces morphological changes in MCF-7 cells as well as erbB-2, erbB-3, and erbB-4 autophosphorylation. MCF-7/ heregulin-transfected cells, which express relatively high levels of HRG, developed estrogen independence and resistance to antiestrogens in vitro and in vivo. This is consistent with a more aggressive hormone-independent phenotype. In contrast with control parental/wild-type cells, estradiol-mediated down-regulation of erbB-2 expression is blocked completely in this particular model system. These results indicate that HRG plays a role in the disruption of ER function. When a transient transfection with an ERE-CAT construct was introduced into these HRG-transfected MCF-7 cells, we observed that the ER was transcriptionally inactive. This suggests that ER signaling is altered in HRG-transfected cells. We observed that overexpression of HRG induces a more aggressive, hormone-independent phenotype that is most likely directly related to the constitutive activation of the erbB-2, erbB-3, and erbB-4 receptor signaling cascade. The data presented here suggest a close cross-regulation between the erbB-2/4 receptors and ER and provide new insights into the mechanism by which breast cancer cells acquire a hormone-independent phenotype.

Epidermal growth factor receptor vIII enhances tumorigenicity in human breast cancer.

Epidermal growth factor receptor vIII (EGFRvIII) is a tumor-specific, ligand-independent, constitutively active variant of the EGFR. Its expression has been detected in gliomas and various other human malignancies. To more fully characterize the function and potential biological role of EGFRvIII in regulating cell proliferation and in tumorigenesis, we transfected EGFRvIII cDNA into a nontumorigenic, interleukin 3 (IL-3)-dependent murine hematopoietic cell line (32D cells). We observed 32D cells expressing high levels of EGFRvIII (32D/EGFRvIII P5) to be capable of abrogating the IL-3-dependent pathway in the absence of ligands. In contrast, the parental cells, 32D/EGFR, 32D/ErbB-4, and 32D/ErbB-2+ErbB-3 cells, all depended on IL-3 or EGF-like ligands for growth. 32D/EGFRvIII P5 cells subjected to long-term culture conditions in the absence of IL-3 revealed further elevation of EGFRvIII expression levels. These results suggested that the IL-3-independent phenotype is mediated by EGFRvIII. The level of expression is a critical driving force for the IL-3-independent phenotype. Dose-response analysis revealed 32D/EGFRvIII cells to require 500-fold higher concentrations (50 ng/ml) of EGF to further stimulate the EGF-mediated proliferation than in the 32D/EGFR cells (100 pg/ml). Similar effects were also observed in beta-cellulin-mediated proliferation. Moreover, 32D cells expressing high levels of EGFRvIII formed large tumors in nude mice, even when no exogenous EGF ligand was administered. In contrast, no tumors grew in mice injected with 32D/EGFR, 32D/ErbB-4, and 32D/ErbB-2+ErbB-3 cells or low-expressing clone 32D/EGFRvIII C2 cells or the parental 32D cells. The changes of the ligand specificity support the notion for an altered conformation of EGFRvIII to reveal an activated ligand-independent oncoprotein with tumorigenic activity analogous to v-erbB. These studies clearly demonstrate that EGFRvIII is capable of transforming a nontumorigenic, IL-3-dependent murine hematopoietic cell line (32D cells) into an IL-3-independent and ligand-independent malignant phenotype in vitro and in vivo. To delineate the biological significance of EGFRvIII in human breast cancer, we expressed EGFRvIII in the MCF-7 human breast cancer cell line. Expression of EGFRvIII in MCF-7 cells produced a constitutively activated EGFRvIII receptor. Expression of EGFRvIII in MCF-7 cells also elevated ErbB-2 phosphorylation, presumably through heterodimerization and cross-talk. These MCF-7/EGFRvIII transfectants exhibited an approximately 3-fold increase in colony formation in 1% serum with no significant effect observed at higher percentages of serum. A similar result was also seen in anchorage-dependent assays. Furthermore, EGFRvIII expression significantly enhanced tumorigenicity of MCF-7 cells in athymic nude mice with P < 0.001. Collectively, these results provide the first evidence that EGFRvIII could play a pivotal role in human breast cancer progression.

Ribozyme-mediated down-regulation of ErbB-4 in estrogen receptor-positive breast cancer cells inhibits proliferation both in vitro and in vivo.

ErbB-4 is a recently discovered member of the class I receptor tyrosine kinase family (ErbB). Little is known about its expression and its importance in human malignancy. To delineate the biological function of ErbB-4 receptors in breast cancer, we used a hammerhead ribozyme strategy to achieve down-regulation of ErbB-4 receptors in various breast cancer cell lines. We observed that down-regulation of ErbB-4 in estrogen receptor-positive (ER+) human breast cancer cell lines (MCF-7 and T47D), which express relatively high levels of ErbB-4, significantly inhibited colony formation. No effects were observed in estrogen receptor-negative (ER-) MDA-MB-453 cells, which express low levels of endogenous ErbB4 and high levels of ErbB-2 and ErbB-3. This occurred despite the fact that fluorescence-activated cell sorter analysis of these latter cells revealed that the expression of the ErbB-4 receptor was completely abrogated by ribozyme treatment. Furthermore, down-regulation of ErbB-4 in T47D and MCF-7 cells significantly inhibited tumor formation in athymic nude mice (P < 0.03 and P < 0.001, respectively). In addition, NRG-stimulated phosphorylation of ErbB-4- and NRG-induced colony formation was significantly reduced in ribozyme-transfected T47D cells. These data provide the first evidence that elevation of ErbB-4 expression plays a role in the proliferation of some ER+ human breast cancer cell lines (T47D and MCF-7) that express high levels of ErbB-4. We have also investigated the expression of ErbB-4 in human primary breast carcinoma specimens, using immunohistochemical staining with an anti-ErbB-4 monoclonal antibody. ErbB-4 expression was found in 60% of the 50 primary breast tumors examined, and high intense immunoreactivity of ErbB-4 was detected in 18% of these primary breast tumors. ErbB-4 receptor expression appeared to correlate with ER+ primary breast tumors. A similar correlation was also observed in the human breast cancer cell lines. These results provide a better understanding of the biological significance of ErbB-4 receptor in breast cancer. Our data suggest that elevation of the ErbB-4 receptor plays a role in ER+ breast cancer cell proliferation. Moreover, ribozyme technology provides a useful tool to delineate the role of a particular gene product.

ErbB-4 ribozymes abolish neuregulin-induced mitogenesis.

The epidermal growth factor-like receptor tyrosine kinase (ErbB) family is frequently overexpressed in a variety of human carcinomas, including breast cancer. To assist in characterizing the role of ErbB-4 in breast cancer, we generated three specific hammerhead ribozymes targeted to the ErbB-4 mRNA. These ribozymes, Rz6, Rz21, and Rz29, efficiently catalyzed the specific cleavage of ErbB-4 message in a cell-free system. We demonstrated that the neuregulin-induced mitogenic effect was abolished in ribozyme Rz29- and Rz6-transfected 32D/ErbB-4 cells. Inhibition of mitogenesis was characterized by ribozyme-mediated down-regulation of ErbB-4 expression. In addition, we provide the first evidence that different threshold levels of ErbB-4 expression and activation correlate with different responses to neuregulin stimulation. High levels of ErbB-4 expression, phosphorylation, and homodimerization are necessary for neuregulin-stimulated, interleukin 3-independent cell proliferation in the 32D/E4 cells. In the case of Rz29-transfected 32D/E4 cells, low levels of ErbB-4 expression allowed neuregulin-induced phosphorylation but were insufficient to couple the activated receptor to cellular signaling. Furthermore, expression of the functional ErbB-4 ribozyme in T47D human breast carcinoma cells led to a down-regulation of endogenous ErbB-4 expression and a reduction of anchorage-independent colony formation. These studies support the use of ErbB-4 ribozymes to define the role of ErbB-4 receptors in human cancers.

Glomangioma of the lung.

An unusual pulmonary tumor was identified on the basis of light and electron microscopic findings as glomangioma. The ultrastructural findings of intracytoplasmic fibrils with dense bodies, electron-dense plaques, pinocytotic vesicles, and basement membranes are consistent with smooth muscle origin. The differential diagnosis between our tumor and other unusual tumors is discussed. The occurrence of a glomangioma in the lung may indicate the existence of pulmonary glomera.

Inhibition of histamine release and phosphatidylcholine metabolism by 5'-deoxy-5'-isobutylthio-3-deazaadenosine.

3-deaza-SIBA, a postulated analogue of S-adenosylhomocysteine, inhibited the IgE-mediated histamine release and blocked both the choline uptake by the rat basophilic leukemia cells and the incorporation of choline into phosphatidylcholine and lysophosphatidylcholine. Unlike inhibitors of methylation which inhibit only dextran- and IgE-mediated reactions, 3-deaza-SIBA also blocked the histamine release from mast cells induced by other secretagogues, such as compound 48/80, ionophore A23187, polymyxin B and ATP. 3-Deaza-SIBA may perturb membrane functions of basophilic leukemia cells and mast cells by inhibiting the biosynthesis of phosphatidylcholine via choline incorporation, the turnover of which is probably required for cellular degranulation and the release of histamine. Previous studies with 3-deaza-SIBA have been mainly interpreted in terms of its structural similarity to S-adenosylhomocysteine and its potential use as an inhibitor of methylation reactions. In light of our present findings, experiments utilizing 3-deaza-SIBA as a biochemical probe have to be carefully interpreted.

Are all adenomatoid tumors adenomatoid mesotheliomas?

The ultrastructure of three histologically typical adenomatoid tumors of the epididymis is presented. Two of the three tumors showed morphologic features of mesothelium similar to those reported in the literature. The third tumor demonstrated a multilayered basal lamina and Weibel-Palade bodies, both of which have been found consistently in sclerosing hemangiomas. Although the ultrastructure of the first two tumors confirms the theory of a mesothelial origin of adenomatoid tumors, the findings of the third tumor indicate that a histologically typical adenomatoid tumor may not necessarily be a mesothelioma and that electron microscopy is indispensable in establishing their nature. These findings also lead us to suggest that the term adenomatoid tumor should remain in use for light microscopic diagnosis, and that the term adenomatoid mesothelioma should be applied only when the mesothelial nature of an adenomatoid tumor is proven by electron microscopy. For those that show ultrastructural evidence of endothelial origin, the term adenomatoid angioma seems to be appropriate and accurate. It is possible that the adenomatoid angioma represents a variant of the histiocytoid hemangioma.

Müllerian adenosarcoma of the uterine cervix.

We present a tumor of the uterine cervix that was composed of histologically benign glandular epithelium and a malignant stromal component, justifying the term müllerian adenosarcoma. Ultrastructurally the stromal cells resemble the endometrial stromal cells, the tumor cells of an endolymphatic stromal myosis of the uterine cervix, and the stromal cells of a stromomyoma of the uterus. In addition there is evidence strongly suggesting a differentiation toward smooth muscle. The patient exhibited no evidence of disease one year after surgery.

An unusual pelvic tumor with benign glandular, sarcomatous, and Wilms' tumor-like components.

We present a pelvic tumor occurring in a 23 year old woman in which a Wilms' tumor-like element was predominant. The presence of a benign glandular component surrounded by small spindle and round malignant cells paralleled that encountered in adenosarcoma of the uterine body and cervix. The possible histogenesis is discussed. Clinically the recurrent tumor showed evidence of a response to chemotherapy. Treatment was complicated by petit mal seizures, for which adriamycin was responsible. The patient exhibited no evidence of disease 22 months after the first operation.

Unusual cystic epithelial choristoma in a celiac lymph node.

With the exception of müllerian-like glandular inclusions in women, reports of nonendometriotic benign glandular inclusions in abdominal lymph nodes are uncommon. We report the case of a 50-year-old woman with an apparently unique, non-müllerian, benign cystic epithelial choristoma in a celiac lymph node found incidentally at cholecystectomy. This case further expands the spectrum of benign lesions that must be differentiated from metastatic adenocarcinoma. Possible histogenesis is discussed.

Duodenal somatostatin-containing tumor with psammoma bodies.

A duodenal tumor was found in a 68-year-old woman undergoing routine cholecystectomy for chronic cholecystitis associated with cholelithiasis. Microscopically, the tumor contained glandular structures and psammoma bodies. Electron microscopic examination demonstrated numerous intracytoplasmic neurosecretory granules consistent with endocrine D cells. Immunoperoxidase staining for somatostatin was positive in the tumor cells. The tumor appeared to be different from the somatostatinomas reported in the literature in that it was nonfunctional, located in the duodenum, and associated with psammoma bodies.

Acute massive hemorrhage from intestinal Crohn disease. Report of seven cases and review of the literature.

Acute massive intestinal bleeding from Crohn disease occurred in 1.4% of 503 patients undergoing treatment at the New York Hospital-Cornell Medical Center over a 43-year period. Of the entire series, 31% had clinically evident rectal bleeding, while 13% had occult bleeding. Combined with previously reported series, onset of massive bleeding was not influenced by age of patient, duration of Crohn disease, use of corticosteroids, or activity of disease. Surgical therapy gave satisfactory results in patients with life-threatening hemorrhage.

Adrenocortical neoplasms. Prognosis and morphology.

The clinical data and morphologic findings in 16 cases of adrenocortical carcinoma were compared with those in 11 cases of surgically removed functional adenomas and 12 cases of nonfunctional adenomas found at autopsy. Histopathologic changes of architectural disarray, pleomorphism, increased mitotic activity, vascular invasion, hemorrhage, or necrosis were generally reliable criteria for diagnosis of malignancy. However, weight was the parameter that most consistently correlated with outcome, since all patients with tumors under 50 Gm. survived and all lesions of 95 Gm. or over proved to be malignant.

Barr bodies in testis with Klinefelter syndrome.

Barr bodies are described in the interstitial cells of testes of a chromatin-positive patient with Klinefelter syndrome. Ultrastructural studies confirm the origin of the Barr body from the nuclear chromatin. Ultrastructurally the interstitial cells showed diminished, smooth endoplasmic reticulum and lipid droplets probably indicating impaired function.

Anaplastic carcinoma of urinary bladder with oat cell features.

Anaplastic carcinoma of the urinary bladder, with oat cell-like components, is rare. We report such a tumor in a fifty-nine-year-old man, who died of rapidly disseminated carcinoma in one year despite postoperative radiotherapy and chemotherapy.

Histoplasmosis of skin and lymph nodes and chronic lymphocytic leukemia.

Nodular skin and cervical lymph node lesions of histoplasmosis, unassociated with systemic symptoms of the infection, developed in a 63-year-old man with untreated chronic lymphocytic leukemia. The histologic patterns in both the skin and lymph node were those of a lymphoproliferative disorder, but Histoplasma organisms were found within a few scattered histiocytes after a careful search. The subtle clinical and pathologic presentation might lead one to overlook the organisms and to believe that the leukemic process had progressed, resulting in potentially dangerous systemic involvement of histoplasmosis and unnecessary chemotherapy for his leukemic process. Complete remission of histoplasmosis was obtained with amphotericin B therapy.