• c-Myc inhibition decreases CIP2A and reduces BCR-ABL1 tyrosine kinase activity in chronic myeloid leukemia.

      Lucas, Claire; Harris, Robert; Giannoudis, Athina; Clark, Richard; University of Liverpool, Royal Liverpool University hospital (Ferrata Storti Foundation, 2015-05-01)
    • Ca

      Wong, Vincent K-W.; Qiu, Congling; Xu, Su-Wei; Law, Betty Yuen Kwan; Zeng, Wu; Wang, Hui; Michelangeli, Francesco; Dias, Ivo Ricardo De Seabra Rodrigues; Qu, Yuan Qing; Chan, Tsz Wai; et al. (2019-05-23)
      Celastrol exhibits anti-arthritic effect in rheumatoid arthritis (RA), but the role of celastrol-mediated Ca mobilization in treatment of RA remains unelucidated. Here, we illustrate the regulatory role of celastrol-induced Ca signalling in synovial fibroblasts of RA patients and adjuvant-induced arthritis (AIA) in rats. Molecular target of celastrol was determined by computational docking, Ca dynamic and functional assays on SERCA. Ca -mediated autophagy in RASFs/RAFLS and the underlying mechanism were verified by quantification of endogenous LC3-II puncta, immunoblotting, and flow cytometry with the Ca chelator (BAPTA/AM) or suitable inhibitors. The anti-arthritic effect of celastrol, autophagy induction and growth rate of synovial fibroblasts in AIA rats were monitored by microCT and immunofluorescence staining. mRNA from joint tissues of AIA rats was isolated for transcriptional analysis of inflammatory genes. The role of Ca in regulating the identified genes was investigated by knockdown of calmodulin, calpains, and calcineurin. Celastrol inhibited SERCA to induce autophagy-dependent cytotoxicity in RASFs/RAFLS via CaMKKβ-AMPK-mTOR pathway and repressed arthritis symptoms in AIA rats. BAPTA/AM hampered the in vitro and in vivo effectiveness of celastrol. Inflammatory- and autoimmunity-associated genes downregulated by celastrol in joint tissues of AIA rat were restored by BAPTA/AM. Knockdown of calmodulin, calpains, and calcineurin in RAFLS confirmed the role of Ca in celastrol-regulated gene expression. Celastrol triggered Ca signalling to induce autophagic cell death in RASFs/RAFLS and ameliorated arthritis in AIA rats mediated by calcium-dependent/-binding proteins facilitating the exploitation of anti-arthritic drugs based on manipulation of Ca signalling. [Abstract copyright: This article is protected by copyright. All rights reserved.]
    • A call to action for climate change research on Caribbean dry forests

      Nelson, Howard P.; Devenish-Nelson, Eleanor S.; Rusk, Bonnie L.; Geary, Matthew; Lawrence, Andrew J.; Department of Biological Sciences, University of Chester; Grenada Dove Conservation Programme, St. Georges, Grenada (Springer, 2018-04-20)
      Tropical dry forest (TDF) is globally one of the most threatened forest types. In the insular Caribbean, limited land area and high population pressure have resulted in the loss of over 60% of TDF, yet local people’s reliance on these systems for ecosystem services is high. Given the sensitivity of TDF to shifts in precipitation regimes and the vulnerability of the Caribbean to climate change, this study examined what is currently known about the impacts of climate change on TDF in the region. A systematic review (n = 89) revealed that only two studies addressed the ecological response of TDF to climate change. Compared to the rapidly increasing knowledge of the effects of climate change on other Caribbean systems and on TDF in the wider neotropics, this paucity is alarming given the value of these forests. We stress the need for long-term monitoring of climate change responses of these critical ecosystems, including phenological and hotspot analyses as priorities.
    • Camera Traps Confirm the Presence of the White-naped Mangabey Cercocebus lunulatus in Cape Three Points Forest Reserve, Western Ghana

      Stanley, Christina R.; Nolan, Ryan; Welsh, Adam; Dempsey, Andrea; Mono, Joseph Cudjoe; Osei, David; Hartley, Matt; Geary, Matthew; University of Chester; West African Primate Conservation Action; The Forestry Commission of Ghana; Zoo and Wildlife Solutions Ltd (IUCN/SSC Primate Specialist Group, 2019)
      The white-naped mangabey Cercocebus lunulatus is severely threatened by logging, mining, and hunting. In the last decade, wild populations have been confirmed in just three forested areas in Ghana and a handful of sites in neighboring Côte d’Ivoire and Burkina Faso. Sightings of this species were recently reported in a fourth area in Ghana, the Cape Three Points Forest Reserve, a forest patch in western Ghana, 60 km from the nearest recorded wild population, which is in the Ankasa Conservation Area. We deployed 14 camera traps across 21 different locations throughout the reserve, with the intention of confirming the presence of this species. Images of the white-naped mangabey were captured at four locations, consolidating recent evidence for a fourth sub-population of this species in Ghana and providing only the second-ever photograph of a wild member of this species in the country. We observed evidence of numerous illegal anthropogenic activities in the reserve, which threaten these mangabeys, and we make recommendations for the protection of the reserve, essential for the conservation of this highly endangered species.
    • Cancerous inhibitor of protein phosphatase 2A (CIP2A) modifies energy metabolism via 5′ AMP-activated protein kinase signalling in malignant cells

      Austin, James A.; orcid: 0000-0002-5384-5221; Jenkins, Rosalind E.; Austin, Gemma M.; Glenn, Mark A.; Dunn, Karen; Scott, Laura; Lucas, Claire M.; Clark, Richard E. (Portland Press Ltd., 2019-08-15)
      Abstract Cancerous inhibitor of protein phosphatase 2A (CIP2A) is an adverse biomarker across many malignancies. Using K562 cells engineered to have high or low CIP2A expression, we show that high CIP2A levels significantly bias cellular energy production towards oxidative phosphorylation (OXPHOS) rather than glycolysis. Mass spectrometric analysis of CIP2A interactors and isobaric tagging for relative and absolute protein quantitation (ITRAQ) experiments identified many associated proteins, several of which co-vary with CIP2A level. Many of these CIP2A associating and co-varying proteins are involved in energy metabolism including OXPHOS, or in 5′ AMP-activated protein kinase (AMPK) signalling, and manipulating AMPK activity mimics the effects of low/high CIP2A on OXPHOS. These effects are dependent on the availability of nutrients, driven by metabolic changes caused by CIP2A. CIP2A level did not affect starvation-induced AMPK phosphorylation of Unc-51 autophagy activating kinase 1 (ULK-1) at Ser555, but autophagy activity correlated with an increase in AMPK activity, to suggest that some AMPK processes are uncoupled by CIP2A, likely via its inhibition of protein phosphatase 2A (PP2A). The data demonstrate that AMPK mediates this novel CIP2A effect on energy generation in malignant cells.
    • Canine mesenchymal stem cells are neurotrophic and angiogenic: An in vitro assessment of their paracrine activity.

      Johnson, William Eustace Basil; Al Delfi, Ibtesam; Aston University, University of Chester, Veterinary Tissue Bank Ltd (Elsevier, 2016-09-19)
      Mesenchymal stem cells (MSCs) have been used in cell replacement therapies for connective tissue damage, but also can stimulate wound healing through paracrine activity. In order to further understand the potential use of MSCs to treat dogs with neurological disorders, this study examined the paracrine action of adipose-derived canine MSCs on neuronal and endothelial cell models. The culture-expanded MSCs exhibited a MSC phenotype according to plastic adherence, cell morphology, CD profiling and differentiation potential along mesenchymal lineages. Treating the SH-SY5Y neuronal cell line with serum-free MSC culture-conditioned medium (MSC CM) significantly increased SH-SY5Y cell proliferation (P <0.01), neurite outgrowth (P = 0.0055) and immunopositivity for the neuronal marker βIII-tubulin (P = 0.0002). Treatment of the EA.hy926 endothelial cell line with MSC CM significantly increased the rate of wound closure in endothelial cell scratch wound assays (P = 0.0409), which was associated with significantly increased endothelial cell proliferation (P <0.05) and migration (P = 0.0001). Furthermore, canine MSC CM induced endothelial tubule formation in EA.hy926 cells in a soluble basement membrane matrix. Hence, this study has demonstrated that adipose-derived canine MSC CM stimulated neuronal and endothelial cells probably through the paracrine activity of MSC-secreted factors. This supports the use of canine MSC transplants or their secreted products in the clinical treatment of dogs with neurological disorders and provides some insight into possible mechanisms of action.
    • Carbohydrate and caffeine improves high intensity running of elite rugby league interchange players during simulated match play

      Clarke, Jon; Highton, Jamie M.; Close, Graeme L.; Twist, Craig; University of Chester; Warrington Wolves RLFC; Liverpool John Moores University (Lippincott, Williams & Wilkins, 2016-11-19)
      Carbohydrate and caffeine improves high intensity running of elite rugby league interchange players during simulated match play
    • Carbohydrate-protein ingestion during self-regulated simulated multiple-sprint sport activity

      Highton, Jamie M.; Nicholas, Ceri; Twist, Craig; Lamb, Kevin L.; University of Chester (2011-09)
    • Cardiac Rehabilitation Delivery Model for Low-Resource Settings: An International Council of Cardiovascular Prevention and Rehabilitation Consensus Statement

      Grace, Sherry L.; Turk-Adawi, Karam I.; Contractor, Aashish; Atrey, Alison; Campbell, Norman R. C.; Derman, Wayne; Ghisi, Gabriela L. M.; Sarkar, Bidyut K.; Yeo, Tee J.; Lopez-Jimenenez, Francisco; et al. (Elsevier, 2016-08-17)
      Cardiovascular disease (CVD) is a global epidemic, which is largely preventable. Cardiac rehabilitation (CR) is demonstrated to be efficacious and cost-effective for secondary prevention in high-income countries. Given its affordability, CR should be more broadly implemented in middle-income countries as well. Hence, the International Council of Cardiovascular Prevention and Rehabilitation (ICCPR) convened a writing panel to recommend strategies to deliver all core CR components in low-resource settings, namely: (1) initial assessment, (2) lifestyle risk factor management (i.e., diet, tobacco, mental health), (3) medical risk factor management (lipids, blood pressure), (4) education for self-management; (5) return to work; and (6) outcome evaluation. Approaches to delivering these components in alternative, arguably lower-cost settings, such as the home, community and primary care, are provided. Recommendations on delivering each of these components where the most-responsible CR provider is a non-physician, such as an allied healthcare professional or community health care worker, are also provided.
    • A Cartesian co-ordinate system for representing the second to fifth metacarpals in the human hand

      Lewis, Stephen J.; University College Chester (Elsevier, 2004-08)
      Purpose The use of hand radiographs has both clinical and anthropometric applications. However, a method for converting standard bony points within the metacarpus to Cartesian co-ordinates does not exist. Methods A simple method for converting standard bony points of the second to fifth metacarpals to Cartesian co-ordinates is described for the first time. Results Using a small set of measurements and treating these with equations of known voracity, this method is accurate and allows the metacarpus to be interro¬gated via a much wider range of geometrical techniques than has so far been available. Conclusions This method allows naked-eye assessments to be supported or re¬placed by metrical evaluations. It is likely to have both clinical and anthropometric uses.
    • Catherine Raisin

      Burek, Cynthia V.; University of Chester (The National Federation of Women's Institutes, 2011)
      This article discusses the life and career of the geologist and educational pioneer Catherine Raison (1855-1945)
    • Catherine Raisin, a role-model professional geologist

      Burek, Cynthia V. (Blackwell, 2003-05)
      This article discusses the life and career of British geologist Catherine Raisin (1855-1945), especially her time teaching at Bedford College (where she was Head of Geography, Head of Botany, and Head of Geology, and became the first woman appointed as Vice-Principal of a college in 1898).
    • CD164 identifies CD4+ T cells highly expressing genes associated with malignancy in Sezary syndrome: the Sezary signature genes, FCRL3, Tox, and miR-214.

      Benoit, Bernice M.; Jariwala, Neha; O'Connor, Geraldine M.; Oetjen, Landon; Whelan, Timothy M.; Werth, Adrienne; Troxel, Andrea B.; Sicard, Helene; Zhu, Lisa; Miller, Christopher; et al. (SpringerLink, 2016-10-20)
      Sézary syndrome (SS), a leukemic variant of cutaneous T-cell lymphoma (CTCL), is associated with a significantly shorter life expectancy compared to skin-restricted mycosis fungoides. Early diagnosis of SS is, therefore, key to achieving enhanced therapeutic responses. However, the lack of a biomarker(s) highly specific for malignant CD4+ T cells in SS patients has been a serious obstacle in making an early diagnosis. We recently demonstrated the high expression of CD164 on CD4+ T cells from Sézary syndrome patients with a wide range of circulating tumor burdens. To further characterize CD164 as a potential biomarker for malignant CD4+ T cells, CD164+ and CD164−CD4+ T cells isolated from patients with high-circulating tumor burden, B2 stage, and medium/low tumor burden, B1–B0 stage, were assessed for the expression of genes reported to differentiate SS from normal controls, and associated with malignancy and poor prognosis. The expression of Sézary signature genes: T plastin, GATA-3, along with FCRL3, Tox, and miR-214, was significantly higher, whereas STAT-4 was lower, in CD164+ compared with CD164−CD4+ T cells. While Tox was highly expressed in both B2 and B1–B0 patients, the expression of Sézary signature genes, FCRL3, and miR-214 was associated predominantly with advanced B2 disease. High expression of CD164 mRNA and protein was also detected in skin from CTCL patients. CD164 was co-expressed with KIR3DL2 on circulating CD4+ T cells from high tumor burden SS patients, further providing strong support for CD164 as a disease relevant surface biomarker.
    • CD271-selected mesenchymal stem cells from adipose tissue enhance cartilage repair and are less angiogenic than plastic adherent mesenchymal stem cells

      Kohli, Nupur; Johnson, William Eustace Basil; Uchida, Kenzo; Aston University, University of Chester, University of Fukui (Nature, 2019-02-28)
      CD271 is a marker of bone marrow MSCs with enhanced differentiation capacity for bone or cartilage repair. However, the nature of CD271+ MSCs from adipose tissue (AT) is less well understood. Here, we investigated the differentiation, wound healing and angiogenic capacity of plastic adherent MSCs (PA MSCs) versus CD271+ MSCs from AT. There was no difference in the extent to which PA MSCs and CD271+ MSCs formed osteoblasts, adipocytes or chondrocytes in vitro. In contrast, CD271+ MSCs transplanted into athymic rats significantly enhanced osteochondral wound healing with reduced vascularisation in the repair tissue compared to PA MSCs and control animals; there was little histological evidence of mature articular cartilage formation in all animals. Conditioned medium from CD271+ MSC cultures was less angiogenic than PA MSC conditioned medium, and had little effect on endothelial cell migration or endothelial tubule formation in vitro. The low angiogenic activity of CD271+ MSCs and improved early stage tissue repair of osteochondral lesions when transplanted, along with a comparable differentiation capacity along mesenchymal lineages when induced, suggests that these selected cells are a better candidate than PA MSCs for the repair of cartilaginous tissue.
    • Celebration of the Centenary of the first female Fellows: Introduction

      Burek, Cynthia V.; Higgs, Bettie M; University of Chester, University College Cork
      Abstract The Geological Society of London was founded in 1807. In May 1919, the first female Fellows were elected to the Society, 112 years after its foundation. This Special Publication celebrates this centenary Eighteen papers have been gathered to highlight recent research, carried out by 24 authors. The publication also builds on stories introduced in a previous Special Publication of the Geological Society, The Role of Women in the History of Geology edited by Burek & Higgs in 2007, the first book to deal solely with this topic, and Burek (2009). It fills in some of the gaps in knowledge with detail that has only recently been uncovered, leading to more in-depth analysis and reporting. The current publication includes more examples from the 20th century, and a small number into the present century, allowing some trends to be identified. The collective work is finding connections previously undocumented and in danger of being lost forever due to the age of the interviewees. The same work also identifies several common challenges that female geoscientists faced, which are still evident in the current investigations. By building on what went before, filling gaps in knowledge and enriching the histories, interesting nuanced insights have emerged.
    • Changes in anthropometry and performance, and their inter-relationships, across three seasons in elite youth rugby league players

      Waldron, Mark; Worsfold, Paul R.; Twist, Craig; Lamb, Kevin L.; University of New England, Australia ; University of Chester ; University of Chester ; University of Chester (Lippincott, Williams & Wilkins, 2014-11-30)
      This study investigated changes in anthropometry and performance, and their inter-relationships, across three consecutive seasons (under-15 to under-17 age group) in elite youth rugby league players. Each player took part in annual anthropometrical and performance assessments, comprising measurements of stature; body mass; limb lengths and circumference; skinfolds, predicted muscle cross-sectional area (CSA); 20 m speed, counter-movement jump height, vertical power and aerobic power. Lean body mass % changed (P < 0.05) between the under-15 (70.9 ± 5.9 %), under-16 (72.0 ± 5.8 %) and the under-17 age groups (74.1 ± 5.7 %). Likewise, predicted quadriceps muscle cross-sectional area (CSA) also changed (P < 0.05) between each age group (under-15 = 120.9 ± 37.8 cm2; under-16 = 133.2 ± 36.0 cm2; under-17 = 154.8 ± 28.3 cm2). Concomitant changes between the under-15 and under-16 group were found for 20 m speed (3.5 ± 0.1 cf. 3.4 ± 0.2 s; P = 0.008) and predicted jumping power (3611.3 ± 327.3 W cf. 4081.5 ± 453.9 W; P = 0.003). Both lean body mass and quadriceps muscle CSA consistently, related to both 20 m sprint time and jumping power, with r-values ranging between -0.39 to –0.63 (20 m sprint time) and 0.55 to 0.75 (jumping power). Our findings demonstrate the importance of gains in lean body mass across later-adolescence that support the ability to generate horizontal speed and predicted vertical power. This information should inform the expectations and subsequent training programs of elite rugby league practitioners.
    • Changes in cervical keratinocyte gene expression associated with integration of human papillomavirus 16

      Alazawi, William; Pett, Mark; Arch, Barbara N.; Scott, Laurie; Freeman, Tom; Stanley, Margaret A.; Coleman, Nicholas; Medical Research Council Cancer Cell Unit, MRC/Hutchison Research Centre, Cambridge ; Department of Pathology, University of Cambridge ; Institute of Public Health, Cambridge ; Medical Research Council Human Genome Mapping Resource Centre, Cambridge ; Medical Research Council Human Genome Mapping Resource Centre, Cambridge ; Department of Pathology, University of Cambridge ; Medical Research Council Cancer Cell Unit, MRC/Hutchison Research Centre, Cambridge/Department of Pathology, University of Cambridge (American Association for Cancer Research, 2002-12-01)
      Episomal integration is a critical event in human papillomavirus (HPV)-related oncogenesis, although little information is currently available concerning the effect of integration on the host transcriptome. Expression microarrays were used to investigate the effect of integration of HPV16 on gene expression in cervical keratinocytes, using the unique cell line model W12. W12 was generated from a cervical low-grade squamous intraepithelial lesion "naturally" infected with HPV16 and at low passage contains approximately 100 HPV16 episomes/cell. With passage in vitro, integration of viral episomes is associated with the development of phenotypic and genomic abnormalities resembling those seen in cervical neoplastic progression in vivo. The Affymetrix U95A oligonucleotide array that contains probes for 12,600 human transcripts was used and 85 genes from a range of host cell pathways that show changes in expression levels after integration of HPV16 were identified. A range of genes not previously described as being involved in cervical neoplastic progression were identified. Interestingly, integration is associated with up-regulation of numerous IFN-responsive genes, in comparison with a baseline of episomally infected cells. These genes include p48, a component of the primary regulator of the IFN response pathway, IFN-stimulated gene factor 3. The physical state of high-risk HPV may substantially influence the response to IFN in infected keratinocytes.
    • Changes in locomotive rates during senior elite rugby league matches

      Sykes, Dave; Twist, Craig; Nicholas, Ceri; Lamb, Kevin L.; University of Chester (Taylor & Francis, 2011-08-05)
      The aim of this study was to quantify the changes in locomotive rates across the duration of senior elite rugby league matches.
    • Changes in selective biomarkers after transurethral resection of the bladder tumour (TURBT), and their association with Non-muscle invasive bladder cancer (NMIBC) recurrence and progression

      Ella-Twongiis, Peter (University of ChesterUniversity of Chester, 2019-10-01)
      Introduction Bladder Cancer (BC) is the 10th most common cancer in the UK, with about 10,000 new cases annually. It affects more men than women (ratio 3:1). Major risk factors include tobacco, chemical carcinogens, schistosomiasis infection and age. About 75-85% of BC are non-muscle invasive (NMIBC), which is associated with high recurrence and progression rates (50-60% within 7-10 years). Currently, diagnosis, treatment and management of BC is via clinical procedures such as transurethral resection of the bladder tumour (TURBT) and endoscopy. Concerning laboratory investigations, there are no routine biomarkers currently available for identifying BC patients at increased risk of developing recurrence and progression. By monitoring changes in selective biomarkers post-TURBT, any sustained changes may be a predictor of cancer recurrence or progression. The main-focus of this research study was to evaluate changes in selective novel biomarkers and their association with recurrence and progression in BC. Materials & Methods In this research, 40 patients (n=40) scheduled for TURBT at the Wrexham Maelor Hospital, North Wales were recruited after written informed consent. Ethical approval for the project was granted via IRAS (REC4: 14/WA/0033). Venous blood samples were taken at baseline (pre-operative) and following TURBT surgery at 1, 3 and 6 months post-operatively. Bladder tumour samples were also taken during TURBT according to standard procedure. Selective biomarkers to assess inflammation, angiogenesis and tumour growth, were measured using commercially available ELISA and BioPlex multiplex assay kits. Tissue immunoreactivity of novel biomarkers were also assessed in BC tissues using immunohistochemistry, with clinical outcome measures being recorded for all patients. Results Significant increases in serum Cluster of differentiation 31 (CD31) (p=0.003) and Stem Cell Factor (SCF) (p=0.032) concentration, as well as trends of increasing concentration of serum basic Fibroblast Growth Factor (bFGF) (p=0.14), Vascular Endothelial Growth Factor Receptor-1 and 2 (VEGFR-1) (p=0.15), VEGFR-2 (p=0.15) and Follistatin (p=0.40) were observed in BC patients up to 6 months post-operative. There were also significant decreases in serum Macrophage Inflammatory Protein -2 (MIP-2) (p=0.001), Platelet Derived Growth Factor (PDGF) (p=0.012), Matrix Metalloproteinase-9 (MMP-9) (p=0.002) and Vascular Endothelial Growth Factor C (VEGF-C) (p=0.04) serum concentration. Trends of decreasing concentration in MMP-2 (p=0.79), MMP-3 (p=0.15), interleukin-6 (IL-6) (p=0.26), interleukin-8 (IL-8) (p=0.15) and tumour necrosis factor-α (TNF-α) (p=0.69) were observed in BC patients up to 6 months post-operative. There was significant immunoreactivity of CD31 (p< 0.001), CD34 (p< 0.001), Human epidermal growth factor receptor-2 (HER-2) (p=0.032), S100P (p< 0.001), Cyclooxygenase-2 (COX-2) (p< 0.001), VEGFR-3 (p< 0.001), SOX-2 (p< 0.001) and thrombomodulin (p=0.010) in bladder tumours. Although recurrence was significantly associated with cancer grade, there was no association with antibody immunoreactivity. Conclusion Findings from the present study may indicate an alternative approach in the monitoring and management of patients with BC. It is proposed that by allowing urological surgeons access to laboratory markers such as MIP-2, MMP-9, PDGF, SCF, HER-2, Thrombomodulin and CD31 (biomarker profile), potentially, in the future, these biomarkers may be used in addition to, or in combination with, currently used scoring systems to predict cancer recurrence and progression. However, verification and validation of these biomarkers are needed using larger cohorts.
    • Changing patterns of drug use in British sport from the 1960s

      Waddington, Ivan; University College Chester (Taylor & Francis, 2005)
      The objective of this paper is systematically to examine evidence relating to the prevalence and the changing patterns of drug use in British sport in the period from the 1960s to the present. There are four major sources of information about the prevalence of drug use among athletes: investigative journalism, including the writings and testimonials of athletes and others involved in sport; formal investigations, which may have legal or quasi-legal powers; surveys; and results from drug testing. The methodological problems associated with these sources of data are discussed. It is concluded that the data suggest that since the 1960s there has been a substantial increase in the use of performance-enhancing drugs by British athletes. More particularly the data suggest that, in athletics, the use of drugs has spread from the heavy throwing events to many other track and field events, and that it has spread from athletics and weightlifting - the sports in which drugs were most frequently used in the 1960s - to many other sports. The use of performance-enhancing drugs has also spread down from the elite to much lower levels, while the use of drugs is now widespread among non-competitive recreational athletes in other sport-related contexts such as gymnasiums.