Lifelong exercise training promotes the remodelling of the immune system and prostate signalome in a rat model of prostate carcinogenesis.

This work aimed to understand how lifelong exercise training promotes the remodelling of the immune system and prostate signalome in a rat model of PCa. Fifty-five male Wistar rats were divided into four groups: control sedentary, control exercised, induced PCa sedentary and induced PCa exercised. Exercised animals were trained in a treadmill for 53 weeks. Pca induction consisted on the sequential administration of flutamide, N-methyl-N-nitrosourea and testosterone propionate implants. Serum concentrations of C-reactive protein (CRP) and tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) were not different among groups. Peripheral levels of γδ T cells were higher in Pca exercised group than in the PCa sedentary group (p < 0.05). Exercise training also induced Oestrogen Receptor (ESR1) upregulation and Mitogen-activated Protein Kinase 13 (MAPK13) downregulation, changed the content of the phosphorylated (at Ser-104) form of this receptor (coded by the gene ESR1) and seemed to increase Erα phosphorylation and activity in exercised PCa rats when compared with sedentary PCa rats. Our data highlight the exercise-induced remodelling of peripheral lymphocyte subpopulations and lymphocyte infiltration in prostate tissue. Moreover, exercise training promotes the remodelling prostate signalome in this rat model of prostate carcinogenesis.

Pharmacological NF-κB inhibition decreases cisplatin chemoresistance in muscle-invasive bladder cancer and reduces cisplatin-induced toxicities.

Most patients with muscle-invasive bladder cancer (MIBC) are not cured with platinum chemotherapy. Up-regulation of nuclear factor kappa light-chain enhancer of activated B cells (NF-κB) is a major mechanism underlying chemoresistance, suggesting that its pharmacological inhibition may increase platinum efficacy. NF-κB signaling was investigated in two patient cohorts. The Cancer Genome Atlas (TCGA) was used to correlate NF-κB signaling and patient survival. The efficacy of cisplatin plus the NF-κB inhibitor dimethylaminoparthenolide (DMAPT) versus cisplatin or DMAPT alone was tested in vitro. Xenografted and immunocompetent MIBC mouse models were studied in vivo. Platinum-naive claudin-low MIBC showed constitutive NF-κB signaling and this was associated with reduced disease-specific survival in TCGA patients. Chemotherapy up-regulated NF-κB signaling and chemoresistance-associated genes, including SPHK1, PLAUR, and SERPINE1. In mice, DMAPT significantly improved the efficacy of cisplatin in both models. The combination preserved body weight, renal function, and morphology, reduced muscle fatigue and IL-6 serum levels, and did not aggravate immuno-hematological toxicity compared with cisplatin alone. These data provide a rationale for combining NF-κB inhibition with platinum-based chemotherapy and conducting a clinical trial in MIBC patients.

In vivo prostate cancer research: Key interspecies prostate anatomical features for translation medicine.

Prostate cancer (PCa) is a prevalent malignancy affecting men worldwide. Animal models play a crucial role in studying PCa pathology and discovering novel approaches to prevent, detect and treat this disease. However, the challenge of translational medicine is the limited reproducibility and inadequate recapitulation of human conditions in animal models. Therefore, a comprehensive understanding of the macroscopic and microscopic anatomy of the prostate gland among distinct animal species is essential for better translating research findings to clinical practice. This review aims to compare and describe the macroscopic and microscopic anatomy of the prostate gland in humans, rats, and dogs, emphasizing the relevant features. Despite the anatomical differences between these species, rats are a valuable model to study human prostate diseases, once they share some features implicated in carcinogenesis in humans. Dogs, on the other hand, are considered the best model for studying PCa due to the development of spontaneous cancer with a higher incidence when compared with other animals and the development of bone metastases. Moreover, the lymphatic system and the sentinel lymph node role and mapping are similar in dogs and humans. However, it is important to recognize that no animal model can directly mimic all aspects of PCa as the human prostate is anatomically different from that of rats and dogs. Therefore, it is essential to analyze and understand the intra- and interspecies variability when translating research findings into clinical practice. This review highlights the importance of a thorough understanding of the anatomical differences between the prostate gland in humans, rats, and dogs when selecting the appropriate animal model for studying PCa.

Femoral Neck Thickness Index as an Indicator of Proximal Femur Bone Modeling.

The alteration in the shape of the femoral neck is an important radiographic sign for scoring canine hip dysplasia (CHD). Previous studies have reported that the femoral neck thickness (FNT) is greater in dogs with hip joint dysplasia, becoming progressively thicker with disease severity. The main objective of this work was to describe a femoral neck thickness index (FNTi) to quantify FNT and to study its association with the degree of CHD using the Fédération Cynologique Internationale (FCI) scheme. A total of 53 dogs (106 hips) were randomly selected for this study. Two examiners performed FNTi estimation to study intra- and inter-examiner reliability and agreement. The paired t-test, the Bland-Altman plots, and the intraclass correlation coefficient showed excellent agreement and reliability between the measurements of the two examiners and the examiners' sessions. All joints were scored in five categories by an experienced examiner according to FCI criteria. The results from examiner 1 were compared between FCI categories. Hips that were assigned an FCI grade of A (n = 19), B (n = 23), C (n = 24), D (n = 24), and E (n = 16) had a mean ± standard deviation FNTi of 0.809 ± 0.024, 0.835 ± 0.044, 0.868 ± 0.022, 0.903 ± 0.033, and 0.923 ± 0.068, respectively (ANOVA, p < 0.05). Therefore, these results show that FNTi is a parameter capable of evaluating proximal femur bone modeling and that it has the potential to enrich conventional CHD scoring criteria if incorporated into a computer-aided diagnosis capable of detecting CHD.

Femoral parallelism: evaluation and impact of variation on canine hip dysplasia assessment.

Adequate radiographic positioning on the X-ray table is paramount for canine hip dysplasia (HD) screening. The aims of this study were to evaluate femoral parallelism on normal ventrodorsal hip extended (VDHE) view and the effect of femoral angulation (FA) on Norberg Angle (NA) and Hip Congruency Index (HCI). The femoral parallelism was evaluated comparing the alignment of the long femoral axis with the long body axis in normal VDHE views and the effect of FA on NA and HCI on repeated VDHE views with different levels of FA. The femoral long axis in normal VDHE views showed a ranged of FA from -4.85° to 5.85°, mean ± standard deviation (SD) of -0.06 ± 2.41°, 95% CI [-4.88, 4.76°]. In the paired views, the mean ± SD femur adduction of 3.69 ± 1.96° led to a statistically significant decrease NA, and HCI, and femur abduction of 2.89 ± 2.12 led to a statistically significant increase in NA and HCI (p < 0.05). The FA differences were also significantly correlated with both NA differences (r = 0.83) and HCI differences (r = 0.44) (p < 0.001). This work describes a methodology that allows evaluation of femoral parallelism in VDHE views and the results suggest that femur abduction yielded more desirable NA and HCI values and adduction impaired NA and HCI values. The positive linear association of FA with NA and HCI allows the use of regression equations to create corrections, to reduce the influence of poor femoral parallelism in the HD scoring.

Appraising Animal Models of Prostate Cancer for Translational Research: Future Directions.

The growing incidence of prostate cancer has prompted a great investment in basic biology and translational studies to develop new therapies. Multiple animal models have been established to study etiological factors, cancer-preventive strategies and the molecular determinants of aggressiveness and metastases. The rat model of prostate cancer induced by chemical carcinogen N-methyl-N-nitrosourea (MNU) and testosterone exposure has become an important tool to study prostatic carcinogenesis and chemopreventive approaches. Over prolonged treatment, this model develops prostatic lesions that closely mimic those observed in human patients. By modifying the experimental conditions, different research groups have been able to induce a vast spectrum of lesions, ranging from early prostatic intraepithelial neoplasia to metastatic cancer. These carefully tuned experimental settings allowed researchers to test lifestyle interventions, and different pharmacological and chemopreventive strategies. However, this model's great flexibility requires careful planning to ensure that the experimental conditions are adequate to obtain the spectrum of lesions intended. The present review addresses such issues, highlighting the value of the rat prostate cancer model and the multiple challenges and opportunities it offers to researchers worldwide.

Long-Term Exposure to Supraphysiological Levels of Testosterone Impacts Rat Submandibular Gland Proteome.

The salivary glands play a central role in the secretion of saliva, whose composition and volume affect oral and overall health. A lesser-explored dimension encompasses the possible changes in salivary gland proteomes in response to fluctuations in sex hormone levels. This study aimed to examine the effects of chronic exposure to testosterone on salivary gland remodeling, particularly focusing on proteomic adaptations. Therefore, male Wistar rats were implanted with subcutaneous testosterone-releasing devices at 14 weeks of age. Their submandibular glands were histologically and molecularly analyzed 47 weeks later. The results underscored a significant increase in gland mass after testosterone exposure, further supported by histologic evidence of granular duct enlargement. Despite increased circulating sex hormones, there was no detectable shift in the tissue levels of estrogen alpha and androgen receptors. GeLC-MS/MS and subsequent bioinformatics identified 308 proteins in the submandibular glands, 12 of which were modulated by testosterone. Of note was the pronounced upregulation of Klk3 and the downregulation of Klk6 and Klk7 after testosterone exposure. Protein-protein interaction analysis with the androgen receptor suggests that Klk3 is a potential target of androgenic signaling, paralleling previous findings in the prostate. This exploratory analysis sheds light on the response of salivary glands to testosterone exposure, providing proteome-level insights into the associated weight and histological changes.

Validation of the Rat Model of Prostate Cancer: Correlating Seminal Vesicle Lesions With Dorsolateral Prostate Lesions.

BACKGROUND/AIM: Lesions in the seminal vesicle are described in the most used protocols for prostate cancer (PCa) induction. This study aimed to characterize the lesions of seminal vesicles associated with a protocol of PCa induction in rats to contribute to better characterization of this model. MATERIALS AND METHODS: Forty-five male Wistar Unilever rats were randomly divided into two control groups: CONT1 (n=10) and CONT2 (n=10); and two PCa-induced groups: IND1 (n=10) and IND2 (n=15), sacrificed at 35 and 61 weeks, respectively. Animals from the induced groups were exposed to a multistep protocol for PCa induction. Animals, seminal vesicles and dorsolateral prostate were weighed. Seminal vesicles and dorsolateral prostate were submitted to histopathological and immunohistochemical analysis. RESULTS: Animals in which PCa was induced had a lower mean body weight when compared with the control animals (p<0.05). The relative mean seminal vesicle weight was higher in groups with PCa when compared with control groups (p<0.05). Although the differences were not statistically significant, animals from the IND2 group developed more lesions than animals from the IND1 and CONT2 groups. It is worth noting that the animals from group IND2 developed papillary adenomas and carcinomas in situ, which were not observed in any other group. Similar to observations in seminal vesicles, animals from group IND2 developed more dorsolateral prostate lesions than animals from the IND1 group (p<0.05). CONCLUSION: We observed that the longer the exposure to testosterone was, the greater was the incidence of preneoplastic and neoplastic lesions in both the seminal vesicle and the prostate, suggesting that testosterone exposure affects the spectrum of developed lesions.

Beneficial Effects of Broccoli (Brassica oleracea var italica) By-products in Diet-induced Obese Mice.

BACKGROUND/AIM: Obesity currently affects the whole world, with greater incidence in high-income countries, with vast economic and social costs. Broccoli harvest generates many by-products equally rich in bioactive compounds with potential anti-obesity effects. This study aimed to evaluate the anti-obesity effects of broccoli by-products flour (BF) in obese mice. MATERIALS AND METHODS: A commercial high-fat diet formulation (representing a Western diet) was used to induce obesity in mice. BF (0.67% or 1.34% weight/weight) was incorporated as a chemoprevention compound into a control and a hypercholesterolemic diet, at two different concentrations, and fed for 14 weeks to C57BL/6J mice. For a therapeutic approach, two groups were fed with the hypercholesterolemic diet for 10 weeks, and then fed with BF-supplemented diets in the last 4 weeks of the study. RESULTS: BF supplementation helped to maintain a lower body weight, reduced adipose tissue accumulation, and enhanced the basal activity of superoxide dismutase and glutathione S-transferase. Although BF supplementation tended to reduce the relative liver weight increased by the Western diet, the differences were not significant. CONCLUSION: BF appears to have a beneficial effect in preventing weight gain and fat accumulation induced by hypercholesterolemic diets.

Acetabular Coverage Area Occupied by the Femoral Head as an Indicator of Hip Congruency.

Accurate radiographic screening evaluation is essential in the genetic control of canine HD, however, the qualitative assessment of hip congruency introduces some subjectivity, leading to excessive variability in scoring. The main objective of this work was to validate a method-Hip Congruency Index (HCI)-capable of objectively measuring the relationship between the acetabulum and the femoral head and associating it with the level of congruency proposed by the Fédération Cynologique Internationale (FCI), with the aim of incorporating it into a computer vision model that classifies HD autonomously. A total of 200 dogs (400 hips) were randomly selected for the study. All radiographs were scored in five categories by an experienced examiner according to FCI criteria. Two examiners performed HCI measurements on 25 hip radiographs to study intra- and inter-examiner reliability and agreement. Additionally, each examiner measured HCI on their half of the study sample (100 dogs), and the results were compared between FCI categories. The paired t-test and the intraclass correlation coefficient (ICC) showed no evidence of a systematic bias, and there was excellent reliability between the measurements of the two examiners and examiners' sessions. Hips that were assigned an FCI grade of A (n = 120), B (n = 157), C (n = 68), D (n = 38) and E (n = 17) had a mean HCI of 0.739 ± 0.044, 0.666 ± 0.052, 0.605 ± 0.055, 0.494 ± 0.070 and 0.374 ± 0.122, respectively (ANOVA, p < 0.01). Therefore, these results show that HCI is a parameter capable of estimating hip congruency and has the potential to enrich conventional HD scoring criteria if incorporated into an artificial intelligence algorithm competent in diagnosing HD.

Early hip laxity screening and later canine hip dysplasia development.

Background and Aim: Passive hip laxity (PHL) is considered the primary risk factor for canine hip dysplasia (HD) and is estimated, in stress hip radiographs, using the distraction index (DI). The study aimed to associate the early PHL using the hip Distractor of University of Trás-os-Montes and Alto Douro (DisUTAD) and the late HD grades. Materials and Methods: A total of 41 dogs (82 hips) were submitted to a follow-up study. First, between 4 and 12 months of age, dogs were radiographed using the DisUTAD hip distractor and were determined the DI for each hip joint. Then, after 12 months of age, dogs were reevaluated for HD using the conventional hip ventrodorsal projection and hips were evaluated for HD using the Fédération Cynologique Internationale (FCI) scoring system. Results: Hips of dogs' in the second examination with FCI grades of A (n=28), B (n=11), C (n=22), and D and E (n=21) had an early DI of 0.32±0.1, 0.38±0.08, 0.50±0.12, and 0.64±0.11, respectively. Statistical analysis using the general linear model univariate, with the DI as dependent variable and the FCI grades, side and sex as fixed factors, and the post hoc Bonferroni correction test showed significant differences among FCI grades (p<0.05). Conclusion: These results show the association between early DI and the late FCI HD grades and the DisUTAD is recommended for the early canine HD diagnosis.

Effect of elbow angle and weight-bearing on the evaluation of joint congruence in dogs.

Joint congruence is relevant for the assessment of elbow dysplasia, a development disorder in dogs. However, quantification of its variation with joint angle and weight-bearing, important factors in quadrupedal locomotion, is lacking. Our objective was to estimate those effects on the alignment of humeroradioulnar joint surfaces. The usual protocol for screening elbow dysplasia, including mediolateral extended and flexed elbow views, was preceded by a weight-bearing lateromedial standing radiographic view, without sedation. Minimum distances were measured at established points of the humeroradioulnar joints classified with a Grade 0 dysplasia. Sixty elbows (30 Portuguese Pointer dogs) were studied. The joint space between the anconeal process and the ulnar trochlear notch increased with flexion, decreasing with weight-bearing (p < .001). The distance between the humeral trochlea and the trochlear notch at the widest point increased with weight-bearing, while it decreased with flexion. Step and distance between the radial head and the base of the medial coronoid process increased with flexion and decreased with weight-bearing. At this point, physiological changes occurred at the humeroulnar joint space. The methodology allowed to quantify the variations on joint spaces with the elbow angle and weight-bearing, important for the interpretation of congruence in diagnosis and treatment of dysplasia. Reference absolute and indexed values obtained allow future comparisons with other breeds and dysplastic dogs. Physiological variations denoted that radioulnar joint became more incongruent under flexion, and more congruent under weight-bearing, while humeroulnar joint became more congruent under flexion, and more incongruent under weight-bearing.

Imaging diagnosis of canine hip dysplasia with and without human exposure to ionizing radiation.

Hip dysplasia (HD) is one of the most common hereditary orthopaedic diseases in dogs, with serious implications for the quality of life of the affected animals. Radiographic screening is essential for the selection of breeding stock in some at-risk breeds, and radiography is also used in the diagnosis of clinical HD cases. A definitive diagnosis of HD is based on radiographic examination, and the most commonly used view is the ventrodorsal hip extended projection, sometimes in combination with various hip stress-based techniques. Radiographic images require high quality positioning and dogs are usually anesthetized and often manually restrained to facilitate optimal positioning. The 'as low as reasonably achievable' (ALARA) principle used in human radioprotection is not always fulfilled in veterinary practice, except in the UK, where human exposure to ionizing radiation in veterinary medicine is strictly regulated. While each dose of ionizing radiation is small, doses accumulate over a lifetime, which can eventually result in substantial radiation exposure. Therefore, manual restraint should be avoided and mechanical immobilization, sedation or general anaesthesia should be used. This review examines the biological effects of human exposure to ionizing radiation and common sources of veterinary exposure. The diagnostic quality of imaging methods for the diagnosis of canine HD is compared between manually restrained and hands-free dog positioning. Hands-free radiographic techniques are available to assess hip laxity, degenerative joint changes and hip osseous structure while preserving image quality, and can be used to select animals for breeding or for the diagnosis of HD.

An overview of the latest in state-of-the-art murine models for prostate cancer.

INTRODUCTION: Prostate cancer (PCa) is a complex, heterogenous and multifocal disease, which is debilitating for patients and often fatal - due to bone metastasis and castration-resistant cancer. The use of murine models that mimic human disease has been crucial in the development of innovative therapies and for better understanding the mechanisms associated with initiation and progression of PCa. AREAS COVERED: This review presents a critical analysis of murine models for the study of PCa, highlighting their strengths, weaknesses and applications. EXPERT OPINION: In animal models, disease may not occur exactly as it does in humans, and sometimes the levels of efficacy that certain treatments obtain in animal models cannot be translated into clinical practice. To choose the most appropriate animal model for each research work, it is crucial to understand the anatomical and physiological differences between the mouse and the human prostate, while it is also important to identify biological similarities and differences between murine and human prostate tumors. Although significant progress has already been made, thanks to many years of research and study, the number of new challenges and obstacles to overcome mean there is a long and difficult road still to travel.

Comparison of Two Distraction Devices for Assessment of Passive Hip Laxity in Dogs.

Canine hip dysplasia is the most common orthopedic developmental condition in the dog and early hip laxity is the main risk factor. The importance of hip laxity in young animals in the development of hip dysplasia is unanimously recognized among researchers and veterinarians due to its medical applicability in terms of disease control and prevention. In the market, there is some certified hip distractors to promote joint laxity. However, the clinical use of some of these distractors complies with a set of usage rules, that can limit its medical application. In this study was compared the technical quality of radiographs and hip distraction using a certified hip distractor (CertD) and Dis-UTAD in 104 dogs (208 joints). The mean pelvic tilting of 1.5 ± 1.6° and 1.5 ± 1.8° were similar when using the CertD and the Dis-UTAD distractors, respectively (P > 0.05). In the CertD sample, the mean hip distraction index (DI) was 0.46 ± 0.17 and in the Dis-UTAD 0.46 ± 0.16; the mean DI differences was 0.001 ± 0.045, resulting in a non-significant paired t-test (P = 0.65) and a significant intraclass correlation coefficient of 0.96, with the 95% lower limit confidence interval of 0.95 (P < 0.05). The statistical power analysis showed a very low distraction index difference effect size. The results suggest that the statistical reproducibility of CertD hip distraction by the Dis-UTAD and the DI mean differences of 0.001 might be considered without clinical importance. The Dis-UTAD might be considered adequate to promote dog hip laxity.

An Overview of Current Alternative Models for Use in the Context of Prostate Cancer Research.

Prostate cancer is one of the most commonly diagnosed cancers worldwide, particularly in elderly populations. To mitigate the expected increase in prostate cancer-related morbidity and mortality as a result of an expanding aged population, safer and more effective therapeutics are required. To this end, plenty of research is focusing on the mechanisms underlying cancer initiation and development, the metastatic process and on the discovery of new therapies. While animal models are used (mainly rats and mice) for the study of prostate cancer, alternative models and methods are increasingly being considered to replace, or at least reduce, the number of animals used in this particular field of research. In this review, we cover some of the alternative models that are currently available for use in the study of prostate cancer, including: mathematical models; 2-D and 3-D cell cultures; microfluidic devices; the chicken egg chorioallantoic membrane-based model; and zebrafish embryo-based models. The main advantages and limitations, as well as some examples of applications, are given for each type of model. According to our analysis, immortalised cell lines are still the most commonly used models in the field of prostate cancer research. However, the use of alternative models for prostate cancer research will likely become more prevalent in the coming years partly because of the increasing societal pressure to reduce the numbers of laboratory animals. In this context, the development and dissemination of effective non-animal alternative models assumes particular relevance and will be instrumental in leveraging their success. Taking these perspectives into account, we believe that technological advances will lead to more effective cell culture systems, namely 3-D cultures or organ-on-a-chip devices, which can be used to replace animal-based models in prostate cancer research.

HPV16 induces penile intraepithelial neoplasia and squamous cell carcinoma in transgenic mice: first mouse model for HPV-related penile cancer.

Penile cancer is an under-studied disease that occurs more commonly in developing countries and 30-50% of cases show high-risk human papillomavirus (HPV) infection. Therapeutic advances are slow, largely due to the absence of animal models for translational research. Here, we report the first mouse model for HPV-related penile cancer. Ten-week-old mice expressing all the HPV16 early genes under control of the cytokeratin 14 (Krt14) gene promoter and matched wild-type controls were exposed topically to dimethylbenz(a)anthracene (DMBA) or vehicle for 16 weeks. At 30 weeks of age, mice were sacrificed for histological analysis. Expression of Ki67, cytokeratin 14, and of the HPV16 oncogenes E6 and E7 was confirmed using immunohistochemistry and quantitative PCR, respectively. HPV16-transgenic mice developed intraepithelial lesions including condylomas and penile intraepithelial neoplasia (PeIN). Lesions expressed cytokeratin 14 and the HPV16 oncogenes E6 and E7 and showed deregulated cell proliferation, demonstrated by Ki67-positive supra-basal cells. HPV16-transgenic mice exposed to DMBA showed increased PeIN incidence and squamous cell carcinoma. Malignant lesions showed varied histological features closely resembling those of HPV-associated human penile cancers. Wild-type mice showed no malignant or pre-malignant lesions even when exposed to DMBA. These observations provide the first experimental evidence to support the etiological role of HPV16 in penile carcinogenesis. Importantly, this is the first mouse model to recapitulate key steps of HPV-related penile carcinogenesis and to reproduce morphological and molecular features of human penile cancer, providing a unique in vivo tool for studying its biology and advancing basic and translational research. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Hands-Free Conventional Radiographic Ventrodorsal Hip Extended View.

Hip dysplasia (HD) is an important hereditary orthopedic disease in the dog associated with osteoarthritis and inadequate welfare for affected animals. The radiographic ventrodorsal hip extended (VDHE) view is used worldwide to select the better animals for breeding. This view normally is performed with manual restraining of the dog to obtain radiographs with acceptable technical quality. The veterinarian exposition to ionizing radiation is inevitable. In this study, the technical quality of VDHE radiographs and hip measurements was compared in 65 dogs radiographed twice, one with the common veterinarian manual restraining and the other obtained using a hind limb holder device, without the veterinarian within the X-ray room. The variables studied were pelvic tilting, patella displacement index, Norberg angle (NA), and subluxation hip category. The results showed a random distribution of right and left pelvic tilting, patella lateral or medial displacement, and hip subluxation categories in both samples (P > 0.05). The holder device positioning showed a better pelvic symmetry (P < 0.05) and a similar patellar displacement (P > 0.05). The mean ± standard deviation of NA was 101.1° ± 6.2° and 100.9° ± 6.1° in the manual and holder device hind limb restraining, respectively (P > 0.05), and the lower limit of 95% confidence interval of intraclass correlation coefficient was >0.75. These results showed statistical reproducibility of NA measurements by the hind limb holder device, and the examiner was protected from exposure to ionizing radiation within the X-ray room.

Association of lifestyle factors with semen quality: A pilot study conducted in men from the Portuguese Trás-os-Montes and Alto Douro region followed in fertility support consultations.

A cross-sectional pilot study was conducted in men followed in fertility consultations, from the portuguese Trás-os-Montes and Alto Douro region, in order to associate several lifestyle factors with the spermatic parameters. Of a total of 522 men, 373 were compared based on the occupational exposure to harmful factors, smoking habits and practice of physical exercise per week, and the other 149 men according to their body mass index (normal weight vs. overweight vs. obesity). In the absence of harmful occupational factors, physical exercise seems to be associated with sperm quality improvement, whether individuals smoke or not. When exposed to harmful environments, non-smokers that practice physical exercise more than two times per week tended to present the best vitality, normal morphology and sperm concentration (p > .05). However, if they smoke, physical exercise seems not enough to enhance the spermatic parameters. The BMI correlated negatively with the spermatic quality, especially with sperm concentration (p < .05). Concerning men that did not present lifestyle risks associated, the motility, midpiece and tail abnormalities, and teratozoospermia index were significantly worse on obese individuals comparing to overweight men (p < .05). Thus, patients should also be recommended to control their weight and to have a BMI under 30 kg/m2 .

HPV16 is sufficient to induce squamous cell carcinoma specifically in the tongue base in transgenic mice.

Head and neck squamous cell carcinomas (HNSCCs) associated with human papillomavirus (HPV) occur specifically in the tonsils and the tongue base, but the reasons for this specificity remain unknown. We studied the distribution of oral and pharyngeal lesions in HPV16-transgenic mice where the expression of all the HPV16 early genes is targeted to keratinising squamous epithelia by the cytokeratin 14 (Krt14) gene promoter. At 30 weeks of age, 100% of mice developed low- and high-grade intraepithelial dysplasia at multiple sites. Twenty per cent of animals developed invasive cancers that remarkably were restricted to the tongue base, in association with the circumvallate papilla. The lesions maintained expression of CK14 (KRT14) and the HPV16 E6 and E7 oncogenes, and displayed deregulated cell proliferation and up-regulation of p16INK4A . Malignant lesions were poorly differentiated and destroyed the tongue musculature. We hypothesised that the tongue base area might contain a transformation zone similar to those observed in the cervix and anus, explaining why HPV-positive cancers target that area specifically. Immunohistochemistry for two transformation zone markers, CK7 (KRT7) and p63 (TP63), revealed a squamocolumnar junction in the terminal duct of von Ebner's gland, composed of CK7+ luminal cells and p63+ basal cells. Dysplastic and invasive lesions retained diffuse p63 expression but only scattered positivity for CK7. Site-specific HPV-induced carcinogenesis in the tongue base may be explained by the presence of a transformation zone in the circumvallate papilla. This mouse model reproduces key morphological and molecular features of HPV-positive HNSCC, providing a unique in vivo tool for basic and translational research. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.