Stellate cell-specific adhesion molecule protocadherin 7 regulates sinusoidal contraction.

BACKGROUND AND AIMS: Sustained inflammation and hepatocyte injury in chronic liver disease activate HSCs to transdifferentiate into fibrogenic, contractile myofibroblasts. We investigated the role of protocadherin 7 (PCDH7), a cadherin family member not previously characterized in the liver, whose expression is restricted to HSCs. APPROACH AND RESULTS: We created a PCDH7 fl/fl mouse line, which was crossed to lecithin retinol acyltransferase-Cre mice to generate HSC-specific PCDH7 knockout animals. HSC contraction in vivo was tested in response to the HSC-selective vasoconstrictor endothelin-1 using intravital multiphoton microscopy. To establish a PCDH7 null HSC line, cells were isolated from PCDH7 fl/fl mice and infected with adenovirus-expressing Cre. Hepatic expression of PCDH7 was strictly restricted to HSCs. Knockout of PCDH7 in vivo abrogated HSC-mediated sinusoidal contraction in response to endothelin-1. In cultured HSCs, loss of PCDH7 markedly attenuated contractility within collagen gels and led to altered gene expression in pathways governing adhesion and vasoregulation. Loss of contractility in PCDH7 knockout cells was impaired Rho-GTPase signaling, as demonstrated by altered gene expression, reduced assembly of F-actin fibers, and loss of focal adhesions. CONCLUSIONS: The stellate cell-specific cadherin, PCDH7, is a novel regulator of HSC contractility whose loss leads to cytoskeletal remodeling and sinusoidal relaxation.

Molecular Markers of Response to Anti-PD1 Therapy in Advanced Hepatocellular Carcinoma.

BACKGROUND & AIMS: Single-agent anti-PD1 checkpoint inhibitors convey outstanding clinical benefits in a small fraction (∼20%) of patients with advanced hepatocellular carcinoma (aHCC) but the molecular mechanisms determining response are unknown. To fill this gap, we herein analyze the molecular and immune traits of aHCC in patients treated with anti-PD1. METHODS: Overall, 111 tumor samples from patients with aHCC were obtained from 13 centers before systemic therapies. We performed molecular analysis and immune deconvolution using whole-genome expression data (n = 83), mutational analysis (n = 72), and histologic evaluation with an endpoint of objective response. RESULTS: Among 83 patients with transcriptomic data, 28 were treated in frontline, whereas 55 patients were treated after tyrosine kinase inhibitors (TKI) either in second or third line. Responders treated in frontline showed upregulated interferon-γ signaling and major histocompatibility complex II-related antigen presentation. We generated an 11-gene signature (IFNAP), capturing these molecular features, which predicts response and survival in patients treated with anti-PD1 in frontline. The signature was validated in a separate cohort of aHCC and >240 patients with other solid cancer types where it also predicted response and survival. Of note, the same signature was unable to predict response in archival tissue of patients treated with frontline TKIs, highlighting the need for fresh biopsies before immunotherapy. CONCLUSION: Interferon signaling and major histocompatibility complex-related genes are key molecular features of HCCs responding to anti-PD1. A novel 11-gene signature predicts response in frontline aHCC, but not in patients pretreated with TKIs. These results must be confirmed in prospective studies and highlights the need for biopsies before immunotherapy to identify biomarkers of response.

Novel microenvironment-based classification of intrahepatic cholangiocarcinoma with therapeutic implications.

OBJECTIVE: The diversity of the tumour microenvironment (TME) of intrahepatic cholangiocarcinoma (iCCA) has not been comprehensively assessed. We aimed to generate a novel molecular iCCA classifier that incorporates elements of the stroma, tumour and immune microenvironment ('STIM' classification). DESIGN: We applied virtual deconvolution to transcriptomic data from ~900 iCCAs, enabling us to devise a novel classification by selecting for the most relevant TME components. Murine models were generated through hydrodynamic tail vein injection and compared with the human disease. RESULTS: iCCA is composed of five robust STIM classes encompassing both inflamed (35%) and non-inflamed profiles (65%). The inflamed classes, named immune classical (~10%) and inflammatory stroma (~25%), differ in oncogenic pathways and extent of desmoplasia, with the inflammatory stroma showing T cell exhaustion, abundant stroma and KRAS mutations (p<0.001). Analysis of cell-cell interactions highlights cancer-associated fibroblast subtypes as potential mediators of immune evasion. Among the non-inflamed classes, the desert-like class (~20%) harbours the lowest immune infiltration with abundant regulatory T cells (p<0.001), whereas the hepatic stem-like class (~35%) is enriched in 'M2-like' macrophages, mutations in IDH1/2 and BAP1, and FGFR2 fusions. The remaining class (tumour classical: ~10%) is defined by cell cycle pathways and poor prognosis. Comparative analysis unveils high similarity between a KRAS/p19 murine model and the inflammatory stroma class (p=0.02). The KRAS-SOS inhibitor, BI3406, sensitises a KRAS-mutant iCCA murine model to anti-PD1 therapy. CONCLUSIONS: We describe a comprehensive TME-based stratification of iCCA. Cross-species analysis establishes murine models that align closely to human iCCA for the preclinical testing of combination strategies.

Inflamed and non-inflamed classes of HCC: a revised immunogenomic classification.

OBJECTIVE: We previously reported a characterisation of the hepatocellular carcinoma (HCC) immune contexture and described an immune-specific class. We now aim to further delineate the immunogenomic classification of HCC to incorporate features that explain responses/resistance to immunotherapy. DESIGN: We performed RNA and whole-exome sequencing, T-cell receptor (TCR)-sequencing, multiplex immunofluorescence and immunohistochemistry in a novel cohort of 240 HCC patients and validated our results in other cohorts comprising 660 patients. RESULTS: Our integrative analysis led to define: (1) the inflamed class of HCC (37%), which includes the previously reported immune subclass (22%) and a new immune-like subclass (15%) with high interferon signalling, cytolytic activity, expression of immune-effector cytokines and a more diverse T-cell repertoire. A 20-gene signature was able to capture ~90% of these tumours and is associated with response to immunotherapy. Proteins identified in liquid biopsies recapitulated the inflamed class with an area under the ROC curve (AUC) of 0.91; (2) The intermediate class, enriched in TP53 mutations (49% vs 29%, p=0.035), and chromosomal losses involving immune-related genes and; (3) the excluded class, enriched in CTNNB1 mutations (93% vs 27%, p<0.001) and PTK2 overexpression due to gene amplification and promoter hypomethylation. CTNNB1 mutations outside the excluded class led to weak activation of the Wnt-ßcatenin pathway or occurred in HCCs dominated by high interferon signalling and type I antigen presenting genes. CONCLUSION: We have characterised the immunogenomic contexture of HCC and defined inflamed and non-inflamed tumours. Two distinct CTNNB1 patterns associated with a differential role in immune evasion are described. These features may help predict immune response in HCC.

Cytokine-like protein 1-induced survival of monocytes suggests a combined strategy targeting MCL1 and MAPK in CMML.

Mouse models of chronic myeloid malignancies suggest that targeting mature cells of the malignant clone disrupts feedback loops that promote disease expansion. Here, we show that in chronic myelomonocytic leukemia (CMML), monocytes that accumulate in the peripheral blood show a decreased propensity to die by apoptosis. BH3 profiling demonstrates their addiction to myeloid cell leukemia-1 (MCL1), which can be targeted with the small molecule inhibitor S63845. RNA sequencing and DNA methylation pattern analysis both point to the implication of the mitogen-activated protein kinase (MAPK) pathway in the resistance of CMML monocytes to death and reveal an autocrine pathway in which the secreted cytokine-like protein 1 (CYTL1) promotes extracellular signal-regulated kinase (ERK) activation through C-C chemokine receptor type 2 (CCR2). Combined MAPK and MCL1 inhibition restores apoptosis of monocytes from patients with CMML and reduces the expansion of patient-derived xenografts in mice. These results show that the combined inhibition of MCL1 and MAPK is a promising approach to slow down CMML progression by inducing leukemic monocyte apoptosis.

Sex steroid receptors in the ovarian follicles of the lizard Sceloporus torquatus.

Estradiol and progesterone have been recognized as important mediators of reproductive events in the female mainly via binding to their receptors. This study aimed to characterize the immunolocalization of the estrogen receptor alfa (ERα), estrogen receptor beta (ERß) and progesterone receptor (PR) in the ovarian follicles of the lizard Sceloporus torquatus. The localization of steroid receptors has a spatio-temporal pattern that depends on the stage of follicular development. The immunostaining intensity of the three receptors was high in the pyriform cells and the cortex of the oocyte of previtellogenic follicles. During the vitellogenic phase, the granulosa and theca immunostaining was intense even with the modification of the follicular layer. In the preovulatory follicles, the receptors were found in yolk and additionally, ERα was also located in the theca. These observations suggest a role for sex steroids in regulating follicular development in lizards, like other vertebrates.

The effects of training based on Nordic hamstring and sprint exercises on measures of physical fitness and hamstring injury prevention in U19 male soccer players.

This study analysed the effects of a training program based on Nordic hamstring and sprint exercises on physical performance and hamstring injuries in young male soccer players. Forty-nine U19 players were randomly assigned to a control (CG; n = 26) or experimental group (EG; n = 23). Linear sprint and with change of direction (COD) were assessed before and after a 14-week training period. Hamstring injuries were collected during the intervention period. Between-groups analysis revealed differences in linear sprint performance (p = 0.012-0.001) in favour of the EG. Pre-to-post performance increased significantly in the EG for 20 m (effect size [ES] = -0.56) and 30 m (ES = -0.62) sprints, but a significant reduction in some COD parameters was observed (ES = 0.45-0.57). In CG, only a significant reduction in COD with dominant leg was found (ES = 0.63). Significant differences in injury burden in favour of the EG was reported such as (27.87 [CG] vs. 3.82 [EG] absence days/1000 h of exposure, rate ratio = 7.30, 95% CI 3.34-15.99). While injury incidence was not different between the EG and CG. These findings suggest that the training program implemented can improve sprint performance and reduce injury burden.

Effects of body mass-based resistance training on measures of physical fitness and musculotendinous injury incidence and burden in U16 male soccer players.

This study examined the effects of body mass-based resistance training (bmRT) on selected measures of physical fitness and injury incidence and burden in soccer players. Forty-six U16 male soccer players were randomly assigned to an control (CG; n = 26) or experimental group (EG; n = 20) Countermovement jump (CMJ), change of direction (CoD) (i.e., 20 m with one CoD), and linear sprint over 30 m were assessed before and after a 15-weeks training. Any type of musculotendinous injury that occurred throughout the intervention period was recorded. Between-group difference was noted at post-test for CMJ (p = 0.008). Pre-to-post training values increased in the EG (effect size [ES] = 1.01) while in the CG no pre-to-post changes were detected (ES = 0.27). No between-group differences at post-test were observed for sprint and CoD tests. Differences in injury burden were reported (33.28 [CG] vs. 9.55 [EG] absence days/1,000 hours exposure, rate ratio = 3.49, 95% CI 2.03-6.00, p < 0.001), but not in injury incidence. A bmRT programis suitable for improving jumping height but not linear and CoD speed performance. Additionally, bmRT may reduce injury burden, and therefore, the severity of musculotendinous injuries in U16 male soccer players.

Immunomodulatory Effects of Lenvatinib Plus Anti-Programmed Cell Death Protein 1 in Mice and Rationale for Patient Enrichment in Hepatocellular Carcinoma.

BACKGROUND AND AIMS: Lenvatinib is an effective drug in advanced HCC. Its combination with the anti-PD1 (programmed cell death protein 1) immune checkpoint inhibitor, pembrolizumab, has generated encouraging results in phase Ib and is currently being tested in phase III trials. Here, we aimed to explore the molecular and immunomodulatory effects of lenvatinib alone or in combination with anti-PD1. APPROACH AND RESULTS: We generated three syngeneic models of HCC in C57BL/6J mice (subcutaneous and orthotopic) and randomized animals to receive placebo, lenvatinib, anti-PD1, or combination treatment. Flow cytometry, transcriptomic, and immunohistochemistry analyses were performed in tumor and blood samples. A gene signature, capturing molecular features associated with the combination therapy, was used to identify a subset of candidates in a cohort of 228 HCC patients who might respond beyond what is expected for monotherapies. In mice, the combination treatment resulted in tumor regression and shorter time to response compared to monotherapies (P < 0.001). Single-agent anti-PD1 induced dendritic and T-cell infiltrates, and lenvatinib reduced the regulatory T cell (Treg) proportion. However, only the combination treatment significantly inhibited immune suppressive signaling, which was associated with the TGFß pathway and induced an immune-active microenvironment (P < 0.05 vs. other therapies). Based on immune-related genomic profiles in human HCC, 22% of patients were identified as potential responders beyond single-agent therapies, with tumors characterized by Treg cell infiltrates, low inflammatory signaling, and VEGFR pathway activation. CONCLUSIONS: Lenvatinib plus anti-PD1 exerted unique immunomodulatory effects through activation of immune pathways, reduction of Treg cell infiltrate, and inhibition of TGFß signaling. A gene signature enabled the identification of ~20% of human HCCs that, although nonresponding to single agents, could benefit from the proposed combination.

The effects of jump training on measures of physical performance, lower extremities injury incidence and burden in highly trained male soccer players.

This study aimed to analyse the effects of a 16-week jump training program on the physical performance and lower extremities injury profile in semi-professional male soccer players. Participants were randomly assigned to the control group (CG; n = 13; age = 21.7 ± 3.6 years) or the experimental group (EG; n = 10; age = 22.3 ± 3.5 years). Countermovement jump (CMJ) height (cm), 30 m linear sprint time (s) with split times at 10 m and 20 m distances, and change of direction speed (CODS; 10 + 10 m with 90° turn) time (s) with turns using the dominant or non-dominant leg, were assessed before and after the intervention. Lower extremity injuries sustained throughout the intervention period were collected. Significant within-group improvements were found in EG in CMJ (p = 0.01; effect size [ES] = 1.03; large). Additionally, between-group difference after intervention was found in CMJ (F = 4.42; p = 0.013) in favour of EG. Injury burden was 194.86 (CG) vs 71.37 (EG) days of absence/1,000 h (RR = 2.73; 95% CI 2.10-3.54; p < 0.001). No other significant within-group or between-group differences were found. In conclusion, compared to regular soccer training, jump training was effective to improve jumping ability and burden in soccer players.

Molecular characterisation of hepatocellular carcinoma in patients with non-alcoholic steatohepatitis.

BACKGROUND AND AIMS: Non-alcoholic steatohepatitis (NASH)-related hepatocellular carcinoma (HCC) is increasing globally, but its molecular features are not well defined. We aimed to identify unique molecular traits characterising NASH-HCC compared to other HCC aetiologies. METHODS: We collected 80 NASH-HCC and 125 NASH samples from 5 institutions. Expression array (n = 53 NASH-HCC; n = 74 NASH) and whole exome sequencing (n = 52 NASH-HCC) data were compared to HCCs of other aetiologies (n = 184). Three NASH-HCC mouse models were analysed by RNA-seq/expression-array (n = 20). Activin A receptor type 2A (ACVR2A) was silenced in HCC cells and proliferation assessed by colorimetric and colony formation assays. RESULTS: Mutational profiling of NASH-HCC tumours revealed TERT promoter (56%), CTNNB1 (28%), TP53 (18%) and ACVR2A (10%) as the most frequently mutated genes. ACVR2A mutation rates were higher in NASH-HCC than in other HCC aetiologies (10% vs. 3%, p <0.05). In vitro, ACVR2A silencing prompted a significant increase in cell proliferation in HCC cells. We identified a novel mutational signature (MutSig-NASH-HCC) significantly associated with NASH-HCC (16% vs. 2% in viral/alcohol-HCC, p = 0.03). Tumour mutational burden was higher in non-cirrhotic than in cirrhotic NASH-HCCs (1.45 vs. 0.94 mutations/megabase; p <0.0017). Compared to other aetiologies of HCC, NASH-HCCs were enriched in bile and fatty acid signalling, oxidative stress and inflammation, and presented a higher fraction of Wnt/TGF-ß proliferation subclass tumours (42% vs. 26%, p = 0.01) and a lower prevalence of the CTNNB1 subclass. Compared to other aetiologies, NASH-HCC showed a significantly higher prevalence of an immunosuppressive cancer field. In 3 murine models of NASH-HCC, key features of human NASH-HCC were preserved. CONCLUSIONS: NASH-HCCs display unique molecular features including higher rates of ACVR2A mutations and the presence of a newly identified mutational signature. LAY SUMMARY: The prevalence of hepatocellular carcinoma (HCC) associated with non-alcoholic steatohepatitis (NASH) is increasing globally, but its molecular traits are not well characterised. In this study, we uncovered higher rates of ACVR2A mutations (10%) - a potential tumour suppressor - and the presence of a novel mutational signature that characterises NASH-related HCC.

Sex steroids are correlated with environmental factors and body condition during the reproductive cycle in females of the lizard Sceloporus torquatus.

Reproduction is regulated by multiple factors that influence physiology and behavior to ensure the continuity of species. However, more work is needed to examine the complex relationships between environmental factors and endocrine transducers that modulate reproductive cycles, particularly in lizards. Here, we aimed to characterize the variation in plasma sex steroid levels in different stages of the reproductive cycle in the lizard Sceloporus torquatus and assess whether sex steroid levels were related to environmental factors (temperature, photoperiod, precipitation, and relative humidity) and body condition. Plasma concentrations of estradiol (E2) and progesterone (P4) from blood samples were quantified by enzyme-linked immunosorbent assay (ELISA) and radioimmunoanalysis (RIA), respectively. Our results indicate that sex steroid concentrations were positively related to follicular development but negatively related to temperature and precipitation. E2 increased as the follicles grew, and its concentrations were highest in the preovulatory phase. P4 showed a similar pattern and persisted during pregnancy. Changes in body condition were non-significant and mainly unrelated to the reproductive stage and plasma sex steroids. Our findings indicate that sex steroids change depending on the season and reproductive stage. We observed high concentrations of E2 and P4 in the late vitellogenic and preovulatory stages, probably because of their role in promoting vitellogenesis and ovulation. Additionally, we observed that follicular development is correlated with temperature and photoperiod. To better understand the mechanisms underlying reproduction, future studies of captive populations where environmental factors can be manipulated are needed.

Molecular classification and therapeutic targets in extrahepatic cholangiocarcinoma.

BACKGROUND & AIMS: Cholangiocarcinoma (CCA), a deadly malignancy of the bile ducts, can be classified based on its anatomical location into either intrahepatic (iCCA) or extrahepatic (eCCA), each with different pathogenesis and clinical management. There is limited understanding of the molecular landscape of eCCA and no targeted therapy with clinical efficacy has been approved. We aimed to provide a molecular classification of eCCA and identify potential targets for molecular therapies. METHODS: An integrative genomic analysis of an international multicenter cohort of 189 eCCA cases was conducted. Genomic analysis included whole-genome expression, targeted DNA-sequencing and immunohistochemistry. Molecular findings were validated in an external set of 181 biliary tract tumors from the ICGC. RESULTS: KRAS (36.7%), TP53 (34.7%), ARID1A (14%) and SMAD4 (10.7%) were the most prevalent mutations, with ∼25% of tumors having a putative actionable genomic alteration according to OncoKB. Transcriptome-based unsupervised clustering helped us define 4 molecular classes of eCCA. Tumors classified within the Metabolic class (19%) showed a hepatocyte-like phenotype with activation of the transcription factor HNF4A and enrichment in gene signatures related to bile acid metabolism. The Proliferation class (23%), more common in patients with distal CCA, was characterized by enrichment of MYC targets, ERBB2 mutations/amplifications and activation of mTOR signaling. The Mesenchymal class (47%) was defined by signatures of epithelial-mesenchymal transition, aberrant TGFß signaling and poor overall survival. Finally, tumors in the Immune class (11%) had a higher lymphocyte infiltration, overexpression of PD-1/PD-L1 and molecular features associated with a better response to immune checkpoint inhibitors. CONCLUSION: An integrative molecular characterization identified distinct subclasses of eCCA. Genomic traits of each class provide the rationale for exploring patient stratification and novel therapeutic approaches. LAY SUMMARY: Targeted therapies have not been approved for the treatment of extrahepatic cholangiocarcinoma. We performed a multi-platform molecular characterization of this tumor in a cohort of 189 patients. These analyses revealed 4 novel transcriptome-based molecular classes of extrahepatic cholangiocarcinoma and identified ∼25% of tumors with actionable genomic alterations, which has potential prognostic and therapeutic implications.

Multilayer intraclonal heterogeneity in chronic myelomonocytic leukemia.

The functional diversity of cells that compose myeloid malignancies, i.e., the respective roles of genetic and epigenetic heterogeneity in this diversity, remains poorly understood. This question is addressed in chronic myelomonocytic leukemia, a myeloid neoplasm in which clinical diversity contrasts with limited genetic heterogeneity. To generate induced pluripotent stem cell clones, we reprogrammed CD34+ cells collected from a patient with a chronic myelomonocytic leukemia in which whole exome sequencing of peripheral blood monocyte DNA had identified 12 gene mutations, including a mutation in KDM6A and two heterozygous mutations in TET2 in the founding clone and a secondary KRAS(G12D) mutation. CD34+ cells from an age-matched healthy donor were also reprogrammed. We captured a part of the genetic heterogeneity observed in the patient, i.e. we analyzed five clones with two genetic backgrounds, without and with the KRAS(G12D) mutation. Hematopoietic differentiation of these clones recapitulated the main features of the patient's disease, including overproduction of granulomonocytes and dysmegakaryopoiesis. These analyses also disclosed significant discrepancies in the behavior of hematopoietic cells derived from induced pluripotent stem cell clones with similar genetic background, correlating with limited epigenetic changes. These analyses suggest that, beyond the coding mutations, several levels of intraclonal heterogeneity may participate in the yet unexplained clinical heterogeneity of the disease.

Recessive Ataxia Differential Diagnosis Algorithm (RADIAL) Versus Specific Niemann-Pick Type C Suspicion Indices: A Retrospective Algorithm Comparison.

Early diagnosis of Niemann-Pick disease type C (NPC) is crucial to slow the progression of neurological manifestations. Different tools were developed to aid diagnosis of NPC, but to date, no study has compared their performance. We aimed to compare the RADIAL algorithm, intended for the differential diagnosis of autosomal recessive cerebellar ataxias (ARCAs) and NPC-specific suspicion indices (SIs). This study was a retrospective analysis of data from 834 patients with molecularly confirmed ARCAs, including 57 NPC cases (RADIAL cohort). We aimed to compare the algorithm performance of RADIAL (Top 1 and Top 3) with that of four SIs (Original, Refined, 2/3 and 2/7) in discriminating NPC cases and non-NPC cases. We also identified ARCAs closely related to NPC as those with low specificity to detect non-NPC cases and described differential and overlapping features with NPC. Overall, excellent sensitivity and specificity (> 0.90) were achieved with both RADIAL and SI tools for NPC cases. The highest sensitivity was attained with the 2/7 SI, Refined SI and Top 3 RADIAL algorithms. Top 1 and Top 3 RADIAL were the most specific tools, followed by the Original SI. The individual comparison of each ARCA revealed that Wilson disease, PLA2G6-associated neurodegeneration, and hypomyelinating leukodystrophy (POLR3A) are frequent NPC false positives (PLA2G6 and POL3A only with the SIs). Both RADIAL and SI diagnostic approaches showed strong discriminatory potential and may be useful screening tools in different clinical contexts.

Estradiol and estrogen receptor α in the mPOA and MeA in dwarf hamster (Phodopus campbelli) fathers.

E2 and its alpha receptor (ERα) have an essential role in the regulation of maternal behavior. In dwarf hamster (Phodopus campbelli), E2 facilitates the display of paternal care, and it is possible that ERα is part of the neuroendocrine mechanisms that regulate this behavior. The aim of this study was to analyze the influence of copulation, cohabitation with the pregnant mate and the presence of the pups on paternal behavior, circulating E2 levels and the presence of ERα in the medial preoptic area (mPOA) and medial amygdala (MeA) in dwarf hamsters. Eight males were mated with intact females (IFs), 8 with tubally ligated females (TLFs) and 8 with ovariectomized females (OFs). In males mated with IFs, paternal behavior tests were performed after copulation, halfway through pregnancy and 24 h after the birth of their pups. Males mated with TLFs were subjected to paternal behavior tests at equivalent periods as the males mated with IFs. In males mated with OFs, paternal behavior tests were performed on days 1, 5 and 10 of cohabitation. After the last paternal behavior tests, blood samples were taken for quantification of E2 by radioimmunoassay (RIA), and the brains were dissected to determine ERα immunoreactivity (ir) in the mPOA and MeA. Fathers mated with IFs had higher serum E2 concentrations and more ERα-ir cells in the mPOA than those of males mated with TLFs and OFs. These results suggest that E2 and its ERα may be associated with paternity in the dwarf hamster.

Testosterone dependent territorial aggression is modulated by cohabitation with a female in male Mongolian gerbils (Meriones unguiculatus).

Most mammal studies on the neuroendocrine mechanisms of territorial aggression have demonstrated that testosterone (T) is required for the display of territorial aggression. However, the relationship between T and aggression is more complex and may be modulated by social factor. The aim of this study was to determine the role of T in territorial aggression in the Mongolian gerbil (Meriones unguiculatus), and the effect of social factors on the modulation of this behavior. The relationship between T and territorial aggression was analyzed using castration and T replacement in two social contexts: male-male and male-female cohabitation. Plasma T concentrations in males of all groups were quantified by radioimmunoassay (RIA). T concentrations were compared using two-way ANOVA. Only sham-castrated and castrated males with T replacement in male-female cohabitation showed aggression, whereas castrated gerbils in the same condition were not aggressive. This indicates that T is the hormone that maintains territorial aggression, but mating is a modulator stimulus. The modulator effect of mating in territorial aggression was associated with an increase in T, but it seems that other mechanisms are involved in the regulation of this behavior, since castrated males with T replacement in the male-male cohabitation did not exhibit aggression, although they had T concentrations as high as these males that received the same treatment, but that cohabited with a female. These results suggest that T is involved in the mechanisms that regulate territorial aggression in the male Mongolian gerbil, and that the cohabitation with a female modulates this behavior.

Tocilizumab in giant cell arteritis: differences between the GiACTA trial and a multicentre series of patients from the clinical practice.

OBJECTIVES: A potential point of concern among clinicians is whether results derived from the clinical trials can be reasonably applied or generalised to a definable group of patients seen in real world. It can be the case of the GiACTA study that is a phase III randomised controlled trial of tocilizumab (TCZ) in giant cell arteritis (GCA). To address this question, we compared the clinical features and the responses to TCZ from the GiACTA trial patients with those from a series of GCA seen in the daily clinical practice. METHODS: Comparative study of clinical features between patients from the GiACTA trial (overall n=251) and those from a multicentre series of real-world GCA patients undergoing TCZ therapy (n=134). The diagnosis of GCA in the GiACTA trial was established by the ACR modified criteria whereas in the series of real-world patients it was made by using the ACR criteria, a positive biopsy of temporal artery or the presence of imaging techniques consistent with large-vessel vasculitis in individuals who presented cranial symptoms of GCA. GiACTA trial patients received subcutaneous TCZ (162 mg every 1 or 2 weeks) whereas those from the clinical practice series were treated using standard IV dose (8 mg/kg/month) or subcutaneous (162 mg/week). RESULTS: Real-life patients undergoing TCZ were older with longer disease duration and higher values of ESR and had received conventional immunosuppressive therapy (mainly methotrexate) more commonly than those included in the GiACTA trial. Despite clinical differences, TCZ was equally effective in both GiACTA trial and clinical practice patients. However, serious infections were more commonly observed in GCA patients recruited from the clinical practice. CONCLUSIONS: Despite clinical differences with patients recruited in clinical trials, data from real-life patients confirm the efficacy of TCZ in GCA.

A non-invasive method for semen collection and evaluation in small and median size lizards.

The aim of this study was to develop a non-invasive procedure to stimulate ejaculation in median and small lizards for semen collection. After semen collection, we applied a battery of tests to assess the motility (wave and progressive), viability (eosin-nigrosin stain), morphology (normal/abnormal), sperm concentration and ejaculate number, seminal volume and colour. We obtained this fluid from all males of the four species of sceloporine lizards (n = 30) and one species of Mexican horned lizards (n = 7). We found that semen from all males had a liquid-like consistency and a milky-white appearance. Ejaculate volume and sperm concentration varied among the males and the species studied. We also observed that although the mobility, viability and normal sperm morphology showed a wide variability, their average value in each species was high. We conclude that "genital papilla pressure" was an efficient and non-invasive semen obtaining method for small- and medium-sized lizards that does not imply damage to donors and could be used in other lizard species.

Neuronal activation associated with paternal and aversive interactions toward pups in the Mongolian gerbils (Meriones unguiculatus).

Approach/avoid model is used to analyze the neural regulation of maternal behavior in the laboratory rat. This model proposes that the medial preoptic area (mPOA) and bed nucleus of stria terminalis (BNST) are brain regions involved in facilitating mechanisms. By contrast, anterior hypothalamic nucleus (AHN), ventromedial hypothalamic nucleus (VMH), and periaqueductal gray participate in the inhibiting mechanisms of neural regulation of maternal behavior. We hypothesized that there are also facilitating and inhibiting mechanisms in the neural regulation of paternal behavior. Here, we determined which neural areas are activated during paternal and aversive interactions with pups in the Mongolian gerbils (Meriones unguiculatus). By testing paternal behavior, we selected 40 males aggressive toward pups and 20 paternal males. These males were organized into six groups of 10 animals in each group: aggressive males that interacted with pups (AGG-pups) or candy (AGG-candy), paternal males that interacted with pups (PAT-pups) or candy (PAT-candy), and males with testosterone (T)-induced paternal behavior that interacted with pups (IPAT-pups) or candy (IPAT-candy). After interacting with pups or candy, the brains were extracted and analyzed for immunoreactivity (ir) with c-fos. Males that interacted with pups had significantly higher c-fos-ir in the mPOA/BNST than males that interacted with candy. Males that displayed aggression had significantly higher c-fos-ir in the AHN, VMH, and periaqueductal gray than aggressive males that interacted with candy. These results suggest that in the neural regulation of paternal behavior in the Mongolian gerbil underlie positive and negative mechanisms as occurs in maternal behavior.