Tumor microenvironment involvement in colorectal cancer progression via Wnt/β-catenin pathway: Providing understanding of the complex mechanisms of chemoresistance

Colorectal cancer (CRC) continues to be one of the main causes of death from cancer because patients progress unfavorably due to resistance to current therapies. Dysregulation of the Wnt/β-catenin pathway plays a fundamental role in the genesis and progression of several types of cancer, including CRC. In many subtypes of CRC, hyperactivation of the β-catenin pathway is associated with mutations of the adenomatous polyposis coli gene. However, it can also be associated with other causes. In recent years, studies of the tumor microenvironment (TME) have demonstrated its importance in the development and progression of CRC. In this tumor nest, several cell types, structures, and biomolecules interact with neoplastic cells to pave the way for the spread of the disease. Cross-communications between tumor cells and the TME are then established primarily through paracrine factors, which trigger the activation of numerous signaling pathways. Crucial advances in the field of oncology have been made in the last decade. This Minireview aims to actualize what is known about the central role of the Wnt/β-catenin pathway in CRC chemoresistance and aggressiveness, focusing on cross-communication between CRC cells and the TME. Through this analysis, our main objective was to increase the understanding of this complex disease considering a more global context. Since many treatments for advanced CRC fail due to mechanisms involving chemoresistance, the data here exposed and analyzed are of great interest for the development of novel and effective therapies.Fil: Novoa Díaz, María Belén. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Martín, María Julia. Universidad Nacional del Sur. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Química del Sur. Universidad Nacional del Sur. Departamento de Química. Instituto de Química del Sur; ArgentinaFil: Gentili, Claudia Rosana. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentin

Age influence on resistance and deformation of the human sutured meniscal horn in the inmediate postoperative period.

To preserve knee function, surgical repair is indicated when a meniscal root disinsertion occurs. However, this surgery has not yet achieved complete recovery of the joint´s natural biomechanics, with the meniscus-suture interface identified as a potentially determining factor. Knowing the deformation and resistance behavior of the sutured meniscal horn and whether these properties are preserved as the patient ages could greatly contribute to improving repair outcomes. Regarding the influence of age on the sutured meniscal horn tissue, in vitro experimentation revealed that meniscal horn specimens older than 75 years old had a more elastic tissue which was less resistant to cut-out than younger menisci at the suture hole area. However, a thickening of the meniscal horns with age, which was also found, leveled out the difference in the force that initiated the tear, as well as in the maximum force borne by the meniscus in the load-to-failure test

Assessment of Surrogate Models for Research on Resistance and Deformation of Repairs of the Human Meniscal Roots: Porcine or Older Human Models?

This study provides data and rationales to aid researchers in the selection of surrogate models for in vitro assessments of surgically repaired meniscal roots. The results are potentially applicable in experimental in vitro investigations aimed at evaluating the performance of surgical repair both in existing approaches and emerging techniques. It could also be of interest for studies seeking to adjust material models of the meniscal tissue around the suture area for incorporation into computational modelsThis research was funded by Junta de Andalucía, Spain, grant number UMA20-FEDERJA-116

Age influence on cut-out resistance of sutured meniscus: An experimental cadaveric study

Surgical treatment of meniscal root detachment is changing from partial meniscectomy to root reinsertion with sutures due to alterations in the knee joint contact biomechanics and early cartilage loss development. Suture fixation techniques of meniscal roots can be grouped into transtibial and in situ fixations, both techniques pierce the meniscal horn to pass a suture thread through the hole and reconnect meniscus to bone. Despite its importance for the survival of the repair, few works focus on determining the resistance of meniscal tissue cut-out by direct thread traction on the suture hole and, as far as we are aware, the influence of age on this resistance has not been yet studied.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Proyecto UMA20-FEDERJA-11

Method for assessing the breaking starting strength of meniscal horns from suture retention tests.

Surgical repair of a detached meniscal root is performed via a transtibial or in situ technique, using sutures to fix the meniscus to restore knee biomechanics. A critical aspect to prevent repair failure is the capacity of the tissue to hold the sutures subjected to forces throughout surgery and the postoperative period. Previously, visual inspection of video-monitored suture retention tests of meniscal horns and other soft tissues, allowed the identification of a breaking starting point that initiates tissue damage, better representing its suture retention capacity than the final failure. In this scenario, it is key to quantifying the Breaking Starting Strength (BSS). Some studies have analyzed horn resistance at the tissue-suture interface, but a robust methodology for computing the BSS has not been described. This work proposes a method to identify the BSS directly from the load curves of suture retention tests. The methodology has been validated using meniscal horns in human and porcine models.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. UMA20-FEDERJA-11

Involvement of parathyroid hormone-related peptide in the aggressive phenotype of colorectal cancer cells

Colorectal cancer (CRC) remains one of the leading causes of mortality from malignant diseases worldwide. In general terms, CRC presents high heterogeneity due to the influence of different genetic and environmental factors; also, the neoplastic cells are strongly influenced by the extracellular matrix and several surrounding cells, known together as the tumor microenvironment (TME). Bidirectional communication takes place between the tumor and the TME through the release of autocrine and paracrine factors. Parathyroid hormone-related peptide (PTHrP) is a cytokine secreted by a wide variety of tissues and is able to regulate several cellular functions both in physiological as well as in pathological processes. It exerts its effects as a paracrine/autocrine factor, although its mode of action is mainly paracrine. It has been shown that this peptide is expressed by several tumors and that the tumor secretion of PTHrP is responsible for the malignant humoral hypercalcemia. Eight years ago, when our research group started studying PTHrP effects in the experimental models derived from intestinal tumors, the literature available at the time addressing the effects of PTHrP on colorectal tumors was limited, and no articles had been published regarding to the paracrine action of PTHrP in CRC cells. Based on this and on our previous findings regarding the role of PTH in CRC cells, our purpose in recent years has been to explore the role of PTHrP in CRC. We analyzed the behavior of CRC cells treated with exogenous PTHrP, focalizing in the study of the following events: Survival, cell cycle progression and proliferation, migration, chemoresistance, tumor-associated angiogenesis, epithelial to mesenchymal transition program and other events also associated with invasion, such us the induction of cancer stem cells features. This work summarizes the major findings obtained by our investigation group using in vitro and in vivo CRC models that evidence the parti cipation of PTHrP in the acquisition of an aggressive phenotype of CRC cells and the molecular mechanisms involved in these processes. Recently, we found that this cytokine induces this malignant behavior not only by its direct action on these intestinal cells but also through its influence on cells derived from TME, promoting a communication between CRC cells and surrounding cells that contributes to the molecular and morphological changes observed in CRC cells. These investigations establish the basis for our next studies in order to address the clinical applicability of our findings. Recognizing the factors and mechanisms that promote invasion in CRC cells, evasion to the cytotoxic effects of current CRC therapies and thus metastasis is decisive for the identification of new markers with the potential to improve early diagnosis and/or to predict prognosis, to predetermine drug resistance and to provide treatment guidelines that include targeted therapies for this disease.Fil: Novoa Díaz, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Carriere, Pedro Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Martín, María Julia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Química del Sur. Universidad Nacional del Sur. Departamento de Química. Instituto de Química del Sur; ArgentinaFil: Calvo, Natalia Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Gentili, Claudia Rosana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentin

Improved tibiofemoral contact restoration after transtibial reinsertion of the anterior root of the lateral meniscus compared to in situ repair: a biomechanical study

Purpose: To compare biomechanical behaviour of the anterior root of the lateral meniscus (ARLM) after a transtibial repair (TTR) and after an in situ repair (ISR), discussing the reasons for the efficacy of the more advantageous technique. Methods: Eight cadaveric human knees were tested at flexion angles from 0° to 90° in four conditions of their ARLM: intact, detached, reinserted using TTR, and reinserted using ISR. Specimens were subjected to 1000 N of compression, and the contact area (CA), mean pressure (MP), and peak pressure (PP) on the tibial cartilage were computed. For the TTR, traction force on the sutures was registered. Results: ARLM detachment significantly altered contact biomechanics, mainly at shallow flexion. After ISR, differences compared to the healthy group persisted (extension, CA 22% smaller (p = 0.012); at 30°, CA 30% smaller (p = 0.012), MP 21%, and PP 32% higher (both p = 0.017); at 60°, CA 28% smaller (p = 0.012), MP 32%, and PP 49% higher (both p = 0.025). With TTR, alterations significantly decreased compared to the injured group, with no statistical differences from the intact ones observed, except for CA at extension (15% decrease, p = 0.012) and at 30° (12% decrease, p = 0.017). The suture tension after TTR, given as mean(SD), was 36.46(11.75)N, 44.32(11.71)N, 40.38(14.93)N, and 43.18(14.89)N for the four tested flexion angles. Conclusions: Alterations caused by ARLM detachment were partially restored with both ISR and TTR, with TTR showing better results on recovering CA, MP, and PP in the immediate postoperative period. The tensile force was far below the value reported to cause meniscal cut-out in porcine models.Funding for open access charge: Universidad de Málaga / CBUA. Funding for open access publishing: Universidad Málaga/CBUA This work was supported by the Ministerio de Ciencia, Inno vación y Universidades (Spain) under grant agreement RTI2018-094339-B-100 and the Consejería de Economia, Conocimiento, Empresas y Universidades de Andalucia, (Spain) under grant agree ment P20-00294

Análisis de la progresión del daño macroscópico en la interfase menisco-sutura tras reparación de la raíz: estudio biomecánico en modelo porcino

En un modelo porcino de la reparación de la raíz posterior del menisco medial sometido a cargas cíclicas representativas de protocolos de rehabilitación habituales en el post-operatorio temprano, no se observa daño macroscópico en la interfase tejido-sutura generada por el orificio de sutura. El desgarro meniscal en ensayo de tracción a rotura se inicia a valores de carga muy superiores a los esperables en el posoperatorio.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Preliminary Study on Artificial versus Animal-Based Feeding Systems for Amblyomma Ticks (Acari: Ixodidae)

Hard ticks pose a threat to animal and human health. Active life stages need to feed on a vertebrate host in order to complete their life cycle. To study processes such as tick-pathogen interactions or drug efficacy and pharmacokinetics, it is necessary to maintain tick colonies under defined laboratory conditions, typically using laboratory animals. The aim of this study was to test a membrane-based artificial feeding system (AFS) applicable for Amblyomma ticks using Amblyomma tonelliae as a biological model. Adult ticks from a laboratory colony were fed in a membrane-based AFS. For comparison, other A. tonelliae adults were fed on calf and rabbit. The proportions of attached (AFS: 76%; calf/rabbit: 100%) and engorged females (AFS: 47.4%; calf/rabbit: 100%) in the AFS were significantly lower compared to animal-based feeding (p = 0.0265). The engorgement weight of in vitro fed ticks (̲ = 658 mg; SD ± 259.80) did not significantly differ from that of ticks fed on animals (p = 0.3272, respectively 0.0947). The proportion of females that oviposited was 100% for all three feeding methods. However, the incubation period of eggs (̲ = 54 days; SD ± 7) was longer in the AFS compared to conventional animal-based feeding (p = 0.0014); ̲ = 45 days; SD ± 2 in the rabbit and (p = 0.0144). ̲ = 48 days; SD ± 2 in the calf). Egg cluster hatching (̲ = 41%; SD ± 44.82) was lower in the AFS than in the other feeding methods (rabbit: ̲ = 74%; SD ± 20; p = 0.0529; calf: ̲ = 81%; SD ± 22; p = 0.0256). Although the attachment, development, and the hatching of AFS ticks were below those from animal-based feeding, the method may be useful in future experiments. Nevertheless, further experiments with a higher number of tick specimens (including immature life stages) and different attractant stimuli are required to confirm the preliminary results of this study and to evaluate the applicability of AFS for Amblyomma ticks as an alternative to animal-based feeding methods

Neospora caninum truncated recombinant proteins formulated with liposomes and CpG-ODNs triggered a humoral immune response in cattle after immunisation and challenge

Abortions caused by Neospora caninum are a serious problem in cattle production and require effective immunoprophylaxis. The objective of this work was to assess the humoral immune response to four recombinant (r) N. caninum antigens in cattle after immunisation and challenge. MIC1 and MIC3 proteins from the micronemes, SRS2 from the surface of tachyzoites, and GRA7 from the dense granules were expressed as truncated recombinant proteins in Escherichia coli. Cationic liposomes (Lip) and CpG oligodeoxynucleotides (CpG-ODNs) were used as adjuvant. Steers were assigned to three groups of six steers each and were inoculated twice subcutaneously, 21 days apart. The rP + Lip + CpG-ODN group received the truncated recombinant proteins rMIC1, rMIC3, rSRS2 and rGRA7 formulated with the adjuvant; the Lip + CpG-ODN group received the adjuvant alone; and the PBS group received sterile phosphate-buffered saline. All steers were subcutaneously challenged with the NC-1 strain of N. caninum 35 days after the second dose of immunisation. Steers from the rP + Lip + CpG-ODN group developed specific IgG, IgG1 and IgG2 against the four recombinant proteins after immunisation. After challenge, IgG against rMIC1 and rMIC3 was detected in rP + Lip + CpG-ODN group and against rSRS2 and rGRA7 in all groups. IgG1 and IgG2 against the four recombinant proteins remained high after challenge in the rP + Lip + CpG-ODN group. Indirect ELISA detected anti-N. caninum antibodies after challenge in all groups, with the highest level of antibodies being detected in the rP + Lip + CpG-ODN group. The recombinant vaccine formulated with rMIC1, rMIC3, rSRS2 and rGRA7 using Lip + CpG-ODN as adjuvant was immunogenic in cattle and the humoral immune response after challenge was enhanced in vaccinated cattle.Fil: Novoa, María Belén. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; ArgentinaFil: Sarli, Macarena. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; ArgentinaFil: Reidel, Ivana Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; ArgentinaFil: Veaute, Carolina Melania Isabel. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Laboratorio de Inmunologia Experimental.; ArgentinaFil: Valentini, Beatriz Susana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; ArgentinaFil: Primo, Maria Evangelina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; Argentin