Compiling forty years of guppy research to investigate the factors contributing to (non)parallel evolution.

Examples of parallel evolution have been crucial for our understanding of adaptation via natural selection. However, strong parallelism is not always observed even in seemingly similar environments where natural selection is expected to favour similar phenotypes. Leveraging this variation in parallelism within well-researched study systems can provide insight into the factors that contribute to variation in adaptive responses. Here we analyse the results of 36 studies reporting 446 average trait values in Trinidadian guppies, Poecilia reticulata, from different predation regimes. We examine how the extent of predator-driven phenotypic parallelism is influenced by six factors: sex, trait type, rearing environment, ecological complexity, evolutionary history, and time since colonization. Analyses show that parallel evolution in guppies is highly variable and weak on average, with only 24.7% of the variation among populations being explained by predation regime. Levels of parallelism appeared to be especially weak for colour traits, and parallelism decreased with increasing complexity of evolutionary history (i.e., when estimates of parallelism from populations within a single drainage were compared to estimates of parallelism from populations pooled between two major drainages). Suggestive - but not significant - trends that warrant further research include interactions between the sexes and different trait categories. Quantifying and accounting for these and other sources of variation among evolutionary 'replicates' can be leveraged to better understand the extent to which seemingly similar environments drive parallel and nonparallel aspects of phenotypic divergence.

Evaluation of olfactory dysfunction persistence after COVID-19: a prospective study.

OBJECTIVE: Alterations of the olfactory function in patients affected by COVID-19 often have an early onset and a variable duration ranging from a few weeks to months. The aim of this study was to evaluate olfactory dysfunction persistence after recovery from COVID-19, and potential related clinical-demographic conditions. PATIENTS AND METHODS: A total of 76 patients recovered from COVID-19 from at least 20 days with olfactory dysfunction during the infection were included in the study. For the subjective evaluation of olfactory function, a visual analogic scale (VAS) was used. The objective evaluation was performed with the use of the Sniffin' Sticks test. RESULTS: Objective assessment of olfactory function revealed that 48 (63.16%) patients were found to be normosmic (TDI ≥ 30.5), 26 (34.21%) were hyposmic (TDI from 30.5 to 16.5) and two (2.63%) were anosmic (TDI ≤ 16.5) at the time of the evaluation. These results did not show a significant difference between subjective and objective tests (p = 0.45). Most patients recovered their sense of smell within the first two months after recovery while a portion (22.2%) still experienced olfactory alterations 4-6 months after SARS-CoV-2 infection. Patients who had not recovered their sense of smell had a significantly longer period of SARS-CoV-2 positivity compared to patients that fully recovered (36.07 ± 7.78 days vs. 29 ± 7.89 days; p = 0.04). CONCLUSIONS: Our results suggest that the duration of the infection negatively correlates with the recovery of olfactory function.

Methods for safety and endpoint ascertainment: identification of adverse events through scrutiny of negatively adjudicated events.

BACKGROUND: The primary goal of phase 2 and 3 clinical trials is to evaluate the safety and effectiveness of therapeutic interventions, and efficient and reproducible ascertainment of important clinical events, either as clinical outcome events (COEs) or adverse events (AEs), is critical. Clinical outcomes require consistency and clinical judgment, so these events are often adjudicated centrally by clinical events classification (CEC) physician reviewers using standardized definitions. In contrast, AEs are reported by sites to the trial coordinating center based on common reporting criteria set by regulatory authorities and trial sponsors. These different requirements have led to the development of separate tracks for COE and AE review. MAIN BODY: Potential COEs that fail to meet standardized definitions for CEC adjudication - i.e. negatively adjudicated events (NAE) - may meet criteria for AEs. Trial oversight practices require the sponsor to process AEs regardless of how the AEs are submitted; therefore, review of NAEs may be necessary to ensure that important AEs do not go unreported. The Duke Clinical Research Institute (DCRI) developed and implemented a process for scrutinizing NAEs to detect potential missed serious AEs. Initial experience with this process across two trials suggests that approximately 0.2% of NAEs are serious unexpected AEs that were not otherwise reported and another 1.5% are serious expected AEs. CONCLUSIONS: Given their infrequent concealment of serious AEs in two large trials assessing cardiovascular outcomes, routine scrutiny of NAEs to identify AEs is not recommended at this time, though it may be useful in some trials and should be carefully considered by the trial team. Closer integration of data across safety surveillance and endpoint adjudication systems may enable scrutiny of NAEs when indicated while limiting complexity associated with this process.

Evaluation of the CD44 isoform v-6 (sCD44var, v6) in the saliva of patients with laryngeal carcinoma and its prognostic role.

BACKGROUND: The soluble fraction of the CD44 protein (solCD44) may constitute a valuable biological marker of Cancer Stem Cells (CSCs), useful for screening/early detection of laryngeal cancer, and for the prognosis. In previous papers, in fact, we have studied the expression of salivary solCD44 in patients with laryngeal tumors, supporting its use for early diagnosis of laryngeal carcinoma with high sensitivity and specificity, also with prognostic role, useful for clinical outcome. OBJECTIVE: The purpose of present study was to verify the levels of solCD44 isoform v6, sCD44var (v6), in saliva samples of patients with laryngeal carcinoma in our tumoral biobank, to evaluate possible correlations with clinical-anamnestic and prognostic data. METHODS: Study design was retrospective. Salivary samples of 66 patients with laryngeal cancer recruited from January 2012 to December 2013 were selected from our tumoral biobank. For each salivary sample was performed the determination of solCD44 and its isoform v6, sCD44var (v6), by ELISA. Qualitative and quantitative results of the test were correlated with clinical and medical history data. For statistical analysis we used the software MedCalc (versione 12.2.1.0). RESULTS: Concentrations of salivary sCD44var v6 were significantly higher according to the size of the primary tumor (T) (p= 0.001), the tumor site glottic or supraglottic-transglottic (p= 0.005) and according to the metastatic lymph node involvement (p= 0005). Furthermore, tumors in advanced disease (stage III-IV) showed values of salivary sCD44var v6 higher than the tumors in early stage, with a statistically significant difference (p= 0.005). CONCLUSIONS: The determination of the levels of salivary solCD44 v6 may represent a promising prognostic test in laryngeal carcinomas.

Rare adult masseteric rhabdomyosarcoma and a review of the literature.

BACKGROUND: Rhabdomyosarcomas (RMSs) are a group of soft-tissue malignant tumors which derive from primitive skeletal muscle tissue that mainly affect children and adolescents. RMSs are very rare in adults, where they are usually located in the extremities. CASE PRESENTATION: A previously healthy 32-year-old male presented at our ENT (ear, nose and throat) outpatient clinic after experiencing a parotid region swelling for 2 months. The patient was treated surgically by excising the mass and by modified radical ipsilateral neck dissection. Histological and immunohistochemical examination indicated masseteric alveolar RMS with lymphatic metastasis. The patient received radiochemotherapy, and he is still alive with no evidence of disease spread 1 year after diagnosis. CONCLUSION: This is the first case of a masseter alveolar RMS to be reported in the literature in a patient older than 25 years; it highlights the broad spectrum of neoplasms that cause parotid region swellings and the importance of considering rare tumors during differential diagnosis.

18FDG uptake in sinonasal inverted papilloma detected by positron emission tomography/computed tomography.

Inverted papilloma (IP) is a benign but locally aggressive sinonasal tumour. Aggressive surgical treatment has thus been traditionally recommended because of the risk of transformation in squamous carcinoma. CT and MRI are used to evaluate bone destruction and soft-tissue extension before surgery but may be ineffective to differentiate an inverted papilloma from squamous cell carcinoma. In recent years, F-18 Fluorodeoxyglucose positron emission tomography ((18)FDG-PET) is widely used as diffuse imaging procedure for diagnosis and followup of malignancy affecting the head and neck district. To evaluate the utility of (18)FDG-PET/CT in the diagnosis of patients with suspicious lesions for IP, twelve patients with suspicious sinonasal inverted papilloma were selected for this study. (18)FDG-PET/CT imaging was performed, and standard uptake value (SUV) was calculated for each patient. SUV(max) was considered as the maximum value measured in the visualized lesions. Seven of the twelve cases exhibited uptake of (18)FFDG with an SUV(max) ranging from 1 to 8.1. Histopathologic diagnosis after surgery confirmed the diagnosis of IP in five cases; all these cases had an SUV(max) > 3. The five cases, which exhibited absence of (18)FDG uptake, had a histological diagnosis of absence of IP.

The usefulness of toluidine staining as a diagnostic tool for precancerous and cancerous oropharyngeal and oral cavity lesions.

Toluidine blue stain is used as a marker to differentiate lesions at high risk of progression in order to improve early diagnosis of oropharyngeal carcinomas. This study focused on 45 oral mucosal lesions in 32 patients (13 female, 19 male). In 9 cases, multiple biopsies were collected. Of the 45 lesions examined, 26 (57.0%) were defined clinically benign, while 19 (42.3%) were defined as suspected lesions (premalignant or malignant). According to the clinical examination, the sensitivity was 53% (16/30) and for toluidine blue staining 96.2% (26/27) (p = 0.0007). The specificity was 80% (12/15) for clinical examination and 77.7% (14/15) for toluidine blue staining (p = 0.79). In conclusion toluidine blue stain has been shown to be a reliable aid when clinical examination is unable to differentiate lesions at high risk of progression and then it improves early diagnosis for oral cavity and oropharyngeal cancer.

Multiple head and neck tumours and their genetic relationship.

Second primary tumours represent one of the major causes of failure in the treatment of head and neck cancer. Advances in early diagnosis and treatment have improved the patient's disease-specific survival. However, the increase in the occurrence of second primary tumours negatively influences the patient's chance of long-term survival. To understand the molecular events underlying the appearance of head and neck multiple tumours, the clinical history has been evaluated in 2 patients both of whom developed 3 primary tumours of the head and neck. To establish the genetic relationship between the different head and neck cancers which had developed in these 2 patients, loss of heterozygosity was investigated using microsatellite markers located on chromosomes 3p, 9p, 11q, 13q, and 17p. These markers were selected as they frequently demonstrate loss of heterozygosity in head and neck cancer. The following markers were used: D3S1234, D3S1300, D9S170, D11S490, and D17S158. Primer sequences were obtained from the genome database for all of these markers. The third tumour that developed in the first patient, 13 years after the primary, showed loss of heterozygosity on chromosome 17p (in the locus for the gene TP53), which was not present in the previous tumours. All tumours in the second patient showed heterozygosity of chromosome 11 at the locus D11S490. These 2 cases show that multiple tumours can be derived from a genetic alteration of a subclone from previous tumours or from an independent preneoplastic cell clone present in the head and neck mucosa.

Mutations and polymorphisms in mitochondrial DNA in head and neck cancer cell lines.

Changes in mitochondrial DNA have been reported in cancer cells. Since little information exists regarding mt DNA mutations in head and neck, the present study focused on ten head and neck cancer cell lines in the attempt to detect alterations in the ND4 gene sequence. DNA was extracted from 10 head and neck squamous cell carcinoma lines from 9 patients. MtDNA sequences were compared in normal and tumour cell line DNA. In ten head and neck squamous cell carcinoma cell lines, 8 somatic mutations and 5 polymorphisms of the mitochondrial gene for ND4 were found. All 5 polymorphisms were silent. Of the 8 somatic mutations, 3 altered the amino acid sequence suggesting a possible effect on enzyme function. The mitochondrial mutations and polymorphisms found demonstrated that these can serve as clonal markers for individual cell lines and demonstrate that the mitochondrial genome remains stable in the cell lines during in vitro culture.

p53 overexpression on the resection margins as a marker of local recurrence in glottic T1a carcinoma.

Glottic carcinoma, in the early stage, may benefit, with excellent results "quoad vitam atque quoad valitudinem", from various modes of treatment--radiotherapy, laser microsurgery and cordectomy being the most common--in definitive cure of the disease. Nevertheless, recurrence, in relation to treatment, oscillates between 4.3-24.1% with laser CO2 surgery, 5.5-32.4% for cordectomy and 5.3-34% for radiotherapy. Prognostic biological markers of recurrence remain to be elucidated, mainly due to the clinical differences in the subjects examined. The present study focused on patients with glottic T1a carcinoma treated with CO2 laser surgery in which correlation between histo-pathological aspects and expression of p53 protein on resection borders were confirmed by onset of local recurrence. Study population comprised 39 patients treated with CO2 laser surgery (January 1985-December 1991) in Clinical Division of Otorhinolarygology, University of Catania. Survival rate, free from recurrence, at 3 and 5 years for this patient group was 87.1% (34/39) and 82% (32/39), respectively. Disease-free survival at 3 and 5 years, was 86.6% in patients with positive resection margins for carcinoma and 87.5% and 79.1%, respectively, for patients with negative resection margins. Survival rate, free from local recurrence, in p53 positive patients, at 3 and 5 years was 78.9% and 68.4%, respectively. In p53 negative patients, survival, free from local recurrence, at 3 and 5 years, was 94.7%. Presence of overexpression of oncoprotein p53 on borders of resection with aspects of dysplasia of various degrees seems, therefore, a marker of high risk of tumour progression and recurrence.

The nasal mucosa of children with nocturnal enuresis before and after treatment with 1-deamino 8-D-arginine vasopressin spray.

Intranasal administration of 1-deamino 8-D-arginine vasopressin (DDVAP) used for treatment of nocturnal enuresis (NE), might be expected to have various effects on the nasal mucosa, e.g. altering the clearance by the mucociliary apparatus. We evaluated two samples (brushes) of epithelial surface cells from the nasal mucosa, one from each nostril, of 18 children (ten males and eight females) with a mean age of 7.7 years (range: 5-13 years) who were affected by primary NE. Samples were taken before and 1 and 6 months after administration of DDVAP spray. No qualitative changes in the epithelial surface cells from nasal mucosa were recognized and only non-statistically significant increases in percentages of goblet, ciliated, basal and unciliated cells at 1 and 6 months after therapy were observed. Thus, it appears that DDVAP spray can be used for at least 6 months in children without apparent risk of damage to the epithelial surface cells from the nasal mucosa.

[Nuclear DNA content in squamous cell carcinoma of the tongue: prognostic significance]. / Significato prognostico del contenuto di DNA nucleare nei carcinomi squamosi della lingua.

The prognosis is poor for advanced (stage III-IV) carcinomas of the tongue. The appearance of metastatic lymph nodes drastically reduces the 2-years survival rate because, in approximately 30% of the cases, metastatic lymph nodes are already present at the time of diagnosis, even in T1, but go undetected. The incidence of lymph nodes metastases has also been related to histological grading. However, this has not proved to be a reliable prognostic parameter, most likely because of the subjectivity of histopathological interpretation. Analysis of the DNA content, in association with other histopathological parameters, has proved to be a useful prognostic marker for other forms of tumors. This could also be the case for carcinomas of the tongue. The present study involved 26 patients who came under observation at the Otorhinolaryngology Clinic of the University of Catania between January 1990 and February 1995. All patients were subject to biopsy sampling and subsequent treatment. According to the UICC classification, then slides were prepared and stained according to Feulgen. A computerized image analysis system (Image-ProPlus) was used to determine the quantity of tumoral nuclear DNA. Analysis of the nuclear DNA content showed that 7 of the 26 carcinomas of the tongue (26.9%) were diploid, 19 (73.1%) aneuploid and of these 7 were also polypoid. In the aneuploid carcinomas the average DI was 1.50 with a range running from 0.70 to 3.1. The correlation of 3-year survival and the clinically studied DNA content showed that 6 of the 11 stage I-II patients were disease-free and had an average tumoral DNA index of 0.95. On the other hand, those cases which showed progression or local recurrences had an average tumoral DI of 1.25. In the group of stage III-IV patients 5 were disease free and showed an average tumoral DNA index of 1.03 while those showing recurrence or progression of the disease had an index of 1.94. The above results appear to confirm that the risk of progression or recurrence is strictly linked to an increase in the average DNA value.

[The role of p53 tumor suppressor gene as prognostic factor in laryngeal squamous cell carcinoma]. / Ruolo dell'oncoproteina p53 nella dignosi precoce e nell'evoluzione dei carcinomi squamosi della testa e del collo.

Mutations in the p53 gene--which codifies anuclear phosphoprotein that acts as a tumor suppressor gene--is the most common genetic alteration in head and neck cancers. The aim of the present study was to investigate the prognostic significance of p53 protein over expression in squamous cell laryngeal carcinoma. To do so we analyzed 31 patients affected by precancerous lesions of the larynx who had undergone multiple biopsy between 1980 and 1995. Twenty-five of these patients later developed laryngeal carcinoma. In this group of patients, 51 biopsies were performed for precancerous lesions (17 hyperplasia, 3 light dysplasia, 23 moderate dysplasia, 8 severe dysplasia) prior to evidence of laryngeal cancer (2.04 biopsies/patient). In the group of patients who did not develop laryngeal cancer, 18 biopsy were performed (2.2 biopsies/patient) and histology revealed: 5 keratosis, 5 light dysplasia, 4 moderate dysplasia and 4 grave dysplasia. Using the immunohistopathological staining technique, 69 formalin-fixed, paraffin-embedded precancerous samples and 25 laryngeal carcinomas were examined for p53 over expression. The monoclonal antibody Pab 1801 was used with the avidinbiotin immunoperoxidase technique; p53 intensity of expression was assessed and correlated with clinical-pathological parameters. Over expression of the p53 protein was found in 56.8% of the precancerous lesions (41% of the hyperplastic lesions, 66% of light dysplastic lesions, 60% of moderate dysplastic lesions and 75% of severe dysplastic lesions) in the group patients who did develop laryngeal cancer and in 22.2% of the precancerous lesions in the group of patients that did not. The transformed lesions showed a strong correlation between intensity of positivity and grade of cellular atypia. Further in 93.3% of the patients with p53 positive precancerous lesions which later developed into laryngeal cancer, p53 over expression was present in the cancerous lesions. There was no significant correlation between p53 immuno reactivity and such clinico pathological tumor parameters as TNM staging and tumorrecurrence. On the other hand, there was a correlation between p53 overexpression and differentiation grading: p53 overexpression was found in 75% of the poorly differentiated tumors, 58.3% of moderately differentiated and 44.4% of well differentiated tumors. The fact that p53 is detected in preneoplastic lesions suggests that p53 gene alteration takes place very early in laryngeal carcinoma and moderate-to-high p53 expression constitutes a high risk of transformation into cancer; on the other hand low expression may reflect reversible changes that can be attributed to the genotoxic effects of tobacco smoking. In conclusion the present data suggest that p53 over expression could be a good prognostic marker in predicting which precancerous laryngeal lesions will progress into cancer.

[Lymph node inflammatory pseudotumor. Description of a case]. / Pseudotumore infiammatorio linfonodale. Descrizione di un caso.

A case of inflammatory pseudotumor of lymph node in a 56-year-old man is described. The awareness of this benign reaction pattern is of great importance for differential diagnosis with many nodal lesions.

Cytogenetic alterations in laryngeal carcinomas.

The purpose of this study was to determine the most frequent chromosomal abnormalities in laryngeal carcinomas. Biopsy specimens of surgical resections from laryngeal squamous cell carcinomas from 15 patients representing different degrees of histologic differentiation were analyzed in short-term culture. Nine of the 15 tumors were hypodiploid with 41 to 45 chromosomes, and four of the 15 tumors were polyploid with more than 50 chromosomes. The most frequent chromosomal alterations we noted included deletion of the short arm of chromosome 3 in 60%, monosomy of chromosome 11 in 30%, and inversions of chromosome 9 and 16 that were present in 20% of the cases.

[Crohn disease in pregnancy. Considerations on and discussion of a clinical case]. / Morbo di Crohn e gravidanza. Considerazioni e discussione su un caso clinico.

Crohn's disease with acute abdomen in pregnancy is described. The Authors evaluate Crohn's disease together with the diagnostic, prognostic and therapeutic problems and relationships between this disease and pregnancy. They conclude as underlining the rarity of Crohn's disease beginning in pregnancy and so the importance of symptoms related to this disease in fertile age woman to make diagnosis and therapy before pregnancy.

In vitro short-term chemosensitivity test in head and neck tumors.

A new method to test the sensitivity of human tumor cells has been developed. A suspension of mechanically dissociated tumor cells is kept in continuous incubation for 24h, in cultures with antineoplastic agents. Drug induced cell cycle perturbations are monitored by flow cytometric computer analysis and DNA distributions of the cells stained with propidium iodide are expressed in percentage. The test is used in 15 head and neck human solid tumors. The drugs tested were: VCR, EpiDx, CDDP, MTX, 5-FU, CPM, BLM. The results obtained reveal that tumor sensitivity varies independently from the stage and malignity grading. Therapeutic combinations are assigned by selecting the drugs on the basis of the individual in vitro response.