Force Transmission Between the Gastrocnemius and Soleus Sub-Tendons of the Achilles Tendon in Rat.

The Achilles tendon (AT) is comprised of three distinct sub-tendons bound together by the inter-subtendon matrix (ISTM). The interactions between sub-tendons will have important implications for AT function. The aim of this study was to investigate the extent to which the ISTM facilitates relative sliding between sub-tendons, and serves as a pathway for force transmission between the gastrocnemius (GAS) and soleus (SOL) sub-tendons of the rat AT. In this study, ATs were harvested from Wistar rats, and the mechanical behavior and composition of the ISTM were explored. To determine force transmission between sub-tendons, the proximal and distal ends of the GAS and SOL sub-tendons were secured, and the forces at each of these locations were measured during proximal loading of the GAS. To determine the ISTM mechanical behavior, only the proximal GAS and distal SOL were secured, and the ISTM was loaded in shear. Finally, for compositional analysis, histological examination assessed the distribution of matrix proteins throughout sub-tendons and the ISTM. The results revealed distinct differences between the forces at the proximal and distal ends of both sub-tendons when proximal loading was applied to the GAS, indicating force transmission between GAS and SOL sub-tendons. Inter-subtendon matrix tests demonstrated an extended initial low stiffness toe region to enable some sub-tendon sliding, coupled with high stiffness linear region such that force transmission between sub-tendons is ensured. Histological data demonstrate an enrichment of collagen III, elastin, lubricin and hyaluronic acid in the ISTM. We conclude that ISTM composition and mechanical behavior are specialized to allow some independent sub-tendon movement, whilst still ensuring capacity for force transmission between the sub-tendons of the AT.

The ceja-1 sequence as a potential new molecular marker for Paracoccidioides brasiliensis infection.

We have developed a two-step PCR assay that amplifies a region of the ceja-1 sequence that is specific for virulent strains of Paracoccidioides brasiliensis. An internal region of the ceja-1 sequence was chosen for designing primers that were utilised in a single tube heminested PCR protocol to amplify DNA from six virulent strains. PCR specificity was determined by the absence of amplified products with genomic DNA from four non-virulent strains of P. brasiliensis and from eight fungal pathogens, one bacterium, two protozoa, one worm and mouse and human genomic DNA (leucocytes). The fact that the PCR product was only obtained with the genetic material from virulent isolates of P. brasiliensis suggested that this partial amplified sequence might be a marker of virulence for this fungus. The diagnostic potential of this PCR was confirmed by the successful amplification of this fragment with genomic DNA obtained in lymph node aspirate from a patient with paracoccidioidomycosis.

Genetic characterization of morphologically variant strains of Paracoccidioides brasiliensis.

Molecular characterization of Paracoccidioides brasiliensis variant strains that had been preserved under mineral oil for decades was carried out by random amplified polymorphic DNA analysis (RAPD). On P. brasiliensis variants in the transitional phase and strains with typical morphology, RAPD produced reproducible polymorphic amplification products that differentiated them. A dendrogram based on the generated RAPD patterns placed the 14 P. brasiliensis strains into five groups with similarity coefficients of 72%. A high correlation between the genotypic and phenotypic characteristics of the strains was observed. A 750 bp-RAPD fragment found only in the wild-type phenotype strains was cloned and sequenced. Genetic similarity analysis using BLASTx suggested that this RAPD marker represents a putative domain of a hypothetical flavin-binding monooxygenase (FMO)-like protein of Neurospora crassa.

Genetic characterization of morphologically variant strains of Paracoccidioides brasiliensis

Molecular characterization of Paracoccidioides brasiliensis variant strains that had been preserved under mineral oil for decades was carried out by random amplified polymorphic DNA analysis (RAPD). On P. brasiliensis variants in the transitional phase and strains with typical morphology, RAPD produced reproducible polymorphic amplification products that differentiated them. A dendrogram based on the generated RAPD patterns placed the 14 P. brasiliensis strains into five groups with similarity coefficients of 72 percent. A high correlation between the genotypic and phenotypic characteristics of the strains was observed. A 750 bp-RAPD fragment found only in the wild-type phenotype strains was cloned and sequenced. Genetic similarity analysis using BLASTx suggested that this RAPD marker represents a putative domain of a hypothetical flavin-binding monooxygenase (FMO)-like protein of Neurospora crassa.