MITO END-3: efficacy of avelumab immunotherapy according to molecular profiling in first-line endometrial cancer therapy.

BACKGROUND: Immunotherapy combined with chemotherapy significantly improves progression-free survival (PFS) compared to first-line chemotherapy alone in advanced endometrial cancer (EC), with a much larger effect size in microsatellite instability-high (MSI-H) cases. New biomarkers might help to select patients who may have benefit among those with a microsatellite-stable (MSS) tumor. PATIENTS AND METHODS: In a pre-planned translational analysis of the MITO END-3 trial, we assessed the significance of genomic abnormalities in patients randomized to standard carboplatin/paclitaxel without or with avelumab. RESULTS: Out of 125 randomized patients, 109 had samples eligible for next-generation sequencing analysis, and 102 had MSI tested. According to The Cancer Genome Atlas (TCGA), there were 29 cases with MSI-H, 26 with MSS TP53 wild type (wt), 47 with MSS TP53 mutated (mut), and 1 case with POLE mutation. Four mutated genes were present in >30% of cases: TP53, PIK3CA, ARID1A, and PTEN. Eleven patients (10%) had a BRCA1/2 mutation (five in MSI-H and six in MSS). High tumor mutational burden (≥10 muts/Mb) was observed in all MSI-H patients, in 4 out of 47 MSS/TP53 mut, and no case in the MSS/TP53 wt category. The effect of avelumab on PFS significantly varied according to TCGA categories, being favorable in MSI-H and worst in MSS/TP53 mut (P interaction = 0.003); a similar non-significant trend was seen in survival analysis. ARID1A and PTEN also showed a statistically significant interaction with treatment effect, which was better in the presence of the mutation (ARID1A P interaction = 0.01; PTEN P interaction = 0.002). CONCLUSION: The MITO END-3 trial results suggest that TP53 mutation is associated with a poor effect of avelumab, while mutations of PTEN and ARID1A are related to a positive effect of the drug in patients with advanced EC.

Alternative academic approaches for testing homologous recombination deficiency in ovarian cancer in the MITO16A/MaNGO-OV2 trial.

BACKGROUND: The detection of homologous recombination deficiency (HRD) can identify patients who are more responsive to platinum and poly ADP ribose polymerase inhibitors (PARPi). MyChoice CDx (Myriad) is the most used HRD test in ovarian cancer (OC). However, some limitations of commercial tests exist, because of the high rate of inconclusive results, costs, and the impossibility of evaluating functional resistance mechanisms. PATIENTS AND METHODS: Two academic genomic tests and a functional assay, the RAD51 foci, were evaluated to detect HRD. One hundred patients with high-grade OC enrolled in the MITO16A/MaNGO-OV2 trial and treated with first-line therapy with carboplatin, paclitaxel, and bevacizumab were analyzed. RESULTS: The failure rate of the two genomic assays was 2%. The sensitivity in detecting HRD when compared with Myriad was 98.1% and 90.6%, respectively. The agreement rate with Myriad was 0.92 and 0.87, with a Cohen's κ coefficient corresponding to 0.84 and 0.74, respectively. For the RAD51 foci assay, the failure rate was 30%. When the test was successful, discordant results for deficient and proficient tumors were observed, and additional HRD patients were identified compared to Myriad; sensitivity was 82.9%, agreement rate was 0.65, and Cohen's κ coefficient was 0.18. The HRD detected by genomic assays and residual tumor at primary surgery and stage was correlated with progression-free survival at multivariate analysis. CONCLUSIONS: Results suggest the feasibility of academic tests for assessing HRD status that show robust concordance with Myriad and correlation with clinical outcome. The contribution of the functional information related to the RAD51 foci test to the genomic data needs further investigation.

Increased early inflammatory response and osteoclastic activity in gingival tissues following conventional osseous resective surgery compared with the fibre retention technique: a pilot study.

BACKGROUND AND OBJECTIVE: The retention of suprabony connective fibres attached into the root cementum during fibre retention osseous resective surgery (FibReORS) results in a more conservative intrasurgical removal of bone, and limits further bone loss and patient morbidity during healing, compared with conventional osseous resective surgery (ORS). This may be a result of the protective effect of preserved connective tissue over the interproximal sites and the lower activation of the inflammatory mechanisms. Thus, the aim of this pilot study was to compare the expression of inflammatory and osteoclastic activity markers in gingival tissues following FibReORS and ORS in the early postsurgical phase. MATERIAL AND METHODS: Twenty-six posterior sextants requiring osseous resective surgery were selected in 13 patients with chronic periodontitis: 13 sextants were randomly assigned to ORS and 13 to FibReORS in a split-mouth design. Gingival biospies were collected during the surgical sessions and at suture removal. Tissue samples were analysed to evaluate the expression of proinflammatory and immunity regulatory mediators (interleukin-1α, C-X-C motif chemokine ligand 5, interferon-γ and tumour necrosis factor-α), cluster of differentiation 14 (CD14; a monocyte/macrophage marker) and TRAP (an osteoclast marker) using immunohistochemical, immunofluorescence and cytofluorimetric analyses, respectively. RESULTS: Postsurgery, a higher number of inflammatory cells and stronger expression of proinflammatory cytokines were observed in the epithelium and connective tissue of ORS gingival samples compared with FibReORS gingival samples (p < 0.001). This was accompanied by increased numbers of CD14-positive and TRAP-positive cells. CONCLUSION: Retention of the supracrestal connective fibres appears to reduce the postsurgical intensity of the host-mediated inflammatory response.

Immunohistochemical study of RhoC GTPase in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a disease with high mortality and morbidity. Metastasis is a significant prognostic factor of the OSCC patients. The Rho GTPases are signaling proteins that controls important cellular processes in various complex mechanisms involved in carcinogenesis. This study aimed to evaluate the expression pattern of RhoC in OSCC protein by immunohistochemistry in situ. Immunohistochemical reactions were performed for RhoC by the method of avitina-biotin-peroxidase activity in samples OSCC: well differentiated (BD, n=6), moderately differentiated (MD, n=24) and poorly differentiated (PD, n=13). The morphometry was taken by QuickScore (percentage and intensity of staining) and only intensity staining. There was no statistical difference (p>0.05) through none of the modes of morphometric analysis between BD, MD and PD. And the RhoC staining was not associated with the histopathologic grading (χ2 = 4.65, p>0.05). However, the morphological evaluation of immunostained for RhoC in cases BD, MD, PD OSCC, regardless of histopathologic grading. These results suggest that there is no correlation between the RhoC immunoexpression and histopathological grading of OSCC.

Review of cerebellopontine angle medulloblastoma.

Cerebellopontine angle (CPA) medulloblastomas (MB) are rare lesions with few cases previously described in the literature. We report two further cases of CPA MB. The patients were a 22-year-old man and a 26-year-old woman with a mass developing in the CPA. The preoperative radiological diagnosis was vestibular schwannoma in the first case and petrosal meningioma in the second case. The patients were operated on through a retrosigmoid approach. The intraoperative findings revealed an intra-axial tumour and the histological diagnosis was classic type of MB in both cases. We review the literature and discuss pathological and radiological features and possible pathogenesis of CPA MB, underlining the necessity to consider MB in the differential diagnosis of CPA lesions.

A prospective study of long-term dysphagia following total laryngectomy.

OBJECTIVE: To evaluate the degree of dysphagia in patients before and after total laryngectomy using the Performance Status Scale for Head and Neck Cancer Patients (PSS) and to determine the effects of post-operative radiation therapy, neck dissection, and pharyngoesophageal spasm on PSS scores. METHODS: We performed a controlled, prospective study at a tertiary referral university hospital. Twenty consecutive patients undergoing laryngectomy were included. Patients were followed for at least two years post-operative, without evidence of local, regional, or distant disease. Only patients with squamous cell carcinoma limited to the endolarynx requiring total laryngectomy with or without elective neck dissection for surgical management of cancer and with no pre-operative treatment were included. Dysphagia was evaluated by PSS prior to surgery and again two years postoperatively with an emphasis on eating in public and normalcy of diet domains. Video fluoroscopic evaluation of swallowing was performed one year after treatment. RESULTS: The relative number of patients with low mean scores in PSS (i.e. usually swallow paste of fluid food in presence of some selected persons or alone; < or = 50) increased after total laryngectomy (p = 0.04). Patients with lower scores reported more frequent spasm of the pharyngoesophageal segment (p = 0.005). Mean scores of both domains decreased after surgery (p < 0.05). CONCLUSION: Eating in public and normalcy of diet scores decreased in 50% of patients after total laryngectomy.

Extra-anatomical hepatic artery reconstruction following post-embolization iatrogenic dissection and arterial anastomotic rupture in two liver transplant recipients.

When hepatic artery reconstruction is required during hepatic transplantation, this is generally performed with donor vessels. We describe two cases requiring a prosthesis. The first case was a 58-year-old man transplanted for cirrhosis complicated by hepatocellular carcinoma. During transplantation, dissection of the celiac trunk occurred due to arterial embolization and the use of the patient's vessels was impossible. An extra-anatomical bypass between the infra-renal aorta and the donor hepatic artery was performed via the interposition of a graft tube. The second case was a 52-year-old man transplanted for cirrhosis complicated by hepatocellular carcinoma. On day 16, a ruptured anastomosis was suspected and the patient underwent emergency revision laparotomy. Arterial revascularisation was performed with an aortohepatic bypass using a synthetic GoreTex((R)) graft. Patient follow-up was uneventful.

Adenylate cyclase/cAMP pathway downmodulation counteracts apoptosis induced by IFN-alpha in human epidermoid cancer cells.

We have reported previously that interferon-alpha (IFN-alpha) induces apoptosis that is counteracted by an epidermal growth factor (EGF) --> Ras --> extracellular signal-regulated kinase (ERK)-dependent survival response in human epidermoid cancer KB cells. We have studied the effects of the cytokine on the cAMP-dependent pathway in these cells. A decrease in the intracellular cAMP levels was recorded in KB cells treated with IFN-alpha, whereas forskolin induced an increase in the production of cAMP that was reduced in the presence of IFN-alpha, suggesting a reduction in the activity of adenylate cyclase (AC) induced by IFN-alpha. These effects were paralleled by significant change in the expression of some AC catalytic subunit(s) and by reduction in the activity of protein kinase A (PKA). 8-Br-cAMP completely antagonized the reduction of PKA activity induced by IFN-alpha, whereas PKA inhibitor KT5720 enhanced the reduction of the enzyme activity induced by IFN-alpha. We have found that IFN-alpha induced a decrease in cAMP response element binding protein (CREB) phosphorylation without changes in its total expression. The concomitant treatment with IFN-alpha and 8-Br-cAMP potentiated and KT5720 counteracted apoptosis induced by IFN-alpha alone. In conclusion, these data suggest that the decrease in AC/cAMP pathway activity is a survival response to the apoptosis induced by IFN-alpha. Therefore, this pathway could represent a target to enhance the antitumor activity of IFN-alpha.

Differentiation of H9c2 cardiomyoblasts: The role of adenylate cyclase system.

The adenylate cyclase (AC)/cAMP/cAMP-dependent protein kinase pathway controls many biological phenomena. The molecular mechanisms by which cAMP induces alternative commitment towards differentiation or proliferation are not still completely known. The differentiation of myoblast cell lines into myocytes/myotubes represents a well-established model of skeletal muscle differentiation. We analyzed the AC/cAMP pathway during terminal differentiation of H9c2 myoblasts. When cultured in low-serum containing medium, H9c2 myoblasts exit the cell cycle and differentiate into myocytes/myotubes. A key step of this process is the expression of myogenin, an essential transcription factor for the terminal differentiation into myocytes. During this phenomenon we observed a decrease in both cAMP levels and AC activity, which suggests a functional negative role of cAMP on the differentiation process of H9c2 cells. 8-Br-cAMP and other cAMP-elevating agents, such as forskolin, IBMX, and isoproterenol, negatively affected skeletal muscle differentiation of H9c2 myoblasts. Both AC activity down-regulation and intracellular cAMP reduction were accompanied by significant variations in the levels of membrane proteins belonging to the AC system (AC catalytic subunit, G(alphai-1), G(alphas)). The functional relationship between intracellular cAMP content and protein levels of AC system is discussed.

A survey of resident selection procedures in oral and maxillofacial surgery.

PURPOSE: This study was conducted to analyze the current procedures used in oral and maxillofacial surgery resident selection, to compare these selection procedures with those used 2 decades ago, to determine whether any differences exist in the selection procedures between 4-year certificate programs and programs that offer formal medical education, and to provide criteria to assist in the counseling of dental students on the application process for oral and maxillofacial surgery residencies. SUBJECTS AND METHODS: Questionnaires were sent to the 106 oral and maxillofacial surgery graduate training programs accredited by the American Dental Association. To provide for a more direct comparison to the study completed in 1976, the current questionnaire was developed, using the original survey as a model. It was divided into 5 sections: general information, information obtained from the formal application and letters of recommendation, the interview, the decision process, and a retrospective view of past decisions. The results were tabulated and the Pearson chi-square test was used to determine statistical significance when comparing the 4-year certificate programs to the programs that offer formal medical education. RESULTS: Seventy-one responses (75.5%) from nonmilitary programs were returned and analyzed. Thirty-nine responses represented dual-degree (MD) programs. Factors that were considered very important when judging a candidate's written application included dental school class rank (76.1%), dental school basic science grades (70.4%), and dental school clinical grades (63.4%). Dual-degree programs placed a greater emphasis on predental basic science grades (P < .01) and dental national board scores (P < .05). When asked about prior resident selection, 86.7% of the respondents said they would select 80% of their former residents again. In addition, 89.9% of the respondents were satisfied with their current selection process. CONCLUSIONS: The procedures used to select oral and maxillofacial surgery residents are relatively constant among programs. Although the dual-degree and 4-year certificate programs use the same criteria for resident selection, the dual-degree programs place greater emphasis on predental academic performance and on the results of the national dental boards. Criteria used 22 years ago to select residents are still applicable, but there has been a shift in the importance of some variables.

Treatment of v-Ki-ras-transformed SVC1 cells with low retinoic acid induces malignancy reversion associated with ras p21 down-regulation.

The effect of nontoxic, low concentrations (10(-8) M) of retinoic acid (RA) for a relatively long time (28 days) on a Kirsten ras-virus transformed cell line (Ki-SVC1), derived from the rat seminal vesicle epithelium, was investigated. In these experimental conditions, the cell treatment with RA induced a decrease of the proliferation rate, apoptosis and a marked reduction of both anchorage-independent growth and tumorigenicity. These biological responses were either preceded or associated with important changes in adenylate cyclase/protein kinase C signaling pathways, the activation of important apoptosis-linked genes and a marked decrease of the v-Ki-ras p21 protein. The significance of these findings is discussed.

Latex allergy: a review for the dental professional.

Reactions to latex products are reportedly occurring with an increased frequency both in patients and in health care providers. Natural rubber latex is found in numerous products, and adverse latex reactions range from contact dermatitis to life-threatening anaphylaxis. A complete understanding of the pathophysiology of latex allergy provides the foundation for the proper diagnosis and treatment of patients with latex allergy. This article provides the reader with a review of latex allergy and suggests strategies for the management of patients with latex hypersensitivity.

Expression of the hepatitis B virus surface antigen in mammalian cells using an Epstein-barr-virus-derived vector.

The hepatitis B virus surface antigen (HBsAg) gene, under control of the inducible mouse metallothionein I gene promoter, was inserted in an expression vector based on the Epstein-Barr virus (EBV). This vector was introduced into human cells by DNA transfection and clones were selected for their resistance to hygromycin B. The recombinant EBV vector replicates efficiently as an episome in human cells and approximately six copies per cell were found in one clone of hygromycin-B-resistant cells. These cells produce high levels of HBsAg in the presence of metals. The protein is mainly found in the cell medium, suggesting that the HBsAg is secreted from the cells.

Clinical features and hormonal characteristics in a case of ovarian arrhenoblastoma.

We report a case of a 34-year-old woman affected with ovarian arrhenoblastoma characterized by very high testosterone (T) levels (34.0-60.0 ng/ml; n.v.0.2-0.9) and suppressed gonadotropin levels. The physical examination revealed: severe hirsutism, acne, amenorrhea and other virilization signs. Basal hormonal evaluation also showed a markedly elevated 17-hydroxyprogesterone (17-OHP) and a mild delta 4 Androstenedione (A) and dehydroepiandrosterone sulfate (DHEAs) increase. ACTH test induced only slight changes in androgen secretion. By contrast, dexamethasone test greatly decreased A and DHEAs whereas T levels were only partially suppressed. Moreover, hCG test was clearly stimulatory for T and A. Suppressed gonadotropin levels did not respond to LHRH stimulation. The removal of the neoplasia was followed by normalization of T levels and increase of serum gonadotropins with subsequent restoration of a normal responsiveness to LHRH and resumption of an ovulatory menstrual cycle. This observation suggests that the high T levels played a primary role in the pathogenesis of the gonadotropin suppression and anovulation. Recovery of acne was complete whereas hirsutism score was reduced but still elevated after one year. This may be due to postoperative A and DHEAs levels slightly above the normal range, indicating the presence of adrenal hyperandrogenism.

ras oncogene-induced transformation of a rat seminal vesicle epithelial cell line produces a marked increase of adenylate cyclase and protein kinase C activities.

Cells transformed by Kirsten murine sarcoma virus (Ki-MSV) have basal adenylate cyclase activity (AC) higher than control cells and comparable level of forskolin-stimulated AC activity. Moreover, a higher protein kinase C (PKC) activity was found to be present in the transformed cells. The molecular mechanism underlying the increase of AC activity was investigated. Our findings strongly suggest that this biochemical event is due to a marked decrease of the alpha i negative control of the enzyme, even though the alpha i of transformed cells appears to possess fully functional domains interacting with both the effector enzyme and the agonist-activated receptor.

Role of calcium in chloride secretion mediated by cAMP pathway activation in rabbit distal colon mucosa.

Effects of Ca2+ on adenosine 3',5'-cyclic monophosphate (cAMP)-mediated Cl- secretion were investigated in intact mucosa and isolated crypt cells of rabbit descending colon. Addition of 10 microM prostaglandin (PG)E2 or forskolin to tissues incubated in Ca(2+)-free medium increased the size of short-circuit current (Isc) and Cl- secretion as estimated by unidirectional 36Cl flux measurements (net flux = -2.31 +/- 0.24 vs. -1.22 +/- 0.10 mueq.h-1.cm-2, n = 4, P < 0.001). Addition of 10 microM PGE2 to tissues incubated in 1.2 mM Ca2+ Ringer induced a 7-fold increase in mean cAMP level, whereas it produced an 11-fold increase in tissues exposed to Ca(2+)-free medium. Membrane preparations from whole mucosa incubated in Ca(2+)-free medium displayed a cyclic nucleotide phosphodiesterase activity significantly lower than controls (18.76 +/- 0.54 vs. 31.20 +/- 0.39 pmol cAMP. mg protein-1.min-1, means +/- SE, n = 4, P < 0.001). Ca2+ removal also affected adenylate cyclase (AC) responsiveness to agonists; AC activity increased in controls by 54 and 226% after stimulation with 10 microM PGE2 and forskolin, respectively, but it increased more (77 and 325%, respectively) after incubation in Ca(2+)-free solutions.(ABSTRACT TRUNCATED AT 250 WORDS)

Anoxia-inducible endonuclease activity as a potential basis of the genomic instability of cancer cells.

Normal rat fibroblasts exhibit a staged response to anoxia which in several respects parallels processes activated in malignant tumor cells. We describe here a new element of the anoxic response, the induction by anoxia of a sequestered endonuclease activity. Such activity is elevated approximately 3-fold within anoxic fibroblasts and during Hirt DNA isolation is able to digest chromatin to produce a nucleosomal ladder. However, DNA is not measurably affected within intact cells, and cells retain complete viability as the endonuclease is induced. The anoxia-inducible endonuclease acts without specificity for DNA sequence. Trace leakage of this endonuclease into the nucleus has obvious potential to underlie the known propensity of anoxic cells to undergo amplification and may be associated with the break-related genomic instability of cancer cells.

Protein kinase C activities are increased in rat thyroid epithelial cells expressing v-ras genes.

Both cytoplasmic and membrane-bound protein kinase C activities are increased in: Harvey-Sarcoma Virus, infected thyroid epithelial cells. The cytoplasmic kinase C increase is found in the chromatographic fraction eluted at lower salt concentration (100 mM NaCl-S100), while the more acidic protein fraction eluted at higher salt concentration (35 mM NaCl-S350) is virtually absent. Although the cytoplasmic S100 fraction from the control and ras-virus infected cells display a comparable PBt2 binding activity, they are different in the Ca+2-dependence and the TPA down regulation. In addition, the membranes from the control and ras-virus infected cells are different phosphate acceptors in place of the H1 histones.

Steroid receptors in epithelial ovarian carcinoma: relation to clinical parameters and survival.

Estrogen receptor (ER) and progestin receptor were measured in samples of tumors obtained at first laparotomy from 97 previously untreated patients suffering with a primary ovarian epithelial tumor, for whom a 3-year follow-up was available. The presence or absence of steroid receptors (threshold arbitrarily fixed at 10 fmol/mg of cytoplasmic protein) was determined by the dextran coated charcoal method and related to a number of patient characteristics such as the residual disease (cutoff, 2 cm), histological type, International Federation of Gynecologists and Obstetricians grade and stage, and age. Results were analyzed by univariate and multivariate methods. (a) The tumor ER positivity was associated with better survival; progestin receptor showed a similar trend but did not reach statistical significance. (b) After stratification for residual tumor the association ER positivity/better survival was still statistically significant in the subset of patients with residual tumor greater than 2 cm. (c) When the median survival times were considered it became apparent that progestin receptor absence nullified the effect associated with positive ER. (d) Multivariate analysis confirmed that among the variables considered the main determinants of prognosis were the size of the residual tumor, serous histological type, and positive ER.