Case series on clinical applications of liquid biopsy in pediatric solid tumors: towards improved diagnostics and disease monitoring.

Background And Aims: Solid tumors account for about 30% of all pediatric cancers. The diagnosis is typically based on histological and molecular analysis of a primary tumor biopsy. Liquid biopsies carry several advantages over conventional tissue biopsy.

Intestinal current measurement and nasal potential difference to make a diagnosis of cases with inconclusive genetics and sweat test.

Background: Nasal potential difference (NPD) and intestinal current measurements (ICM) are cystic fibrosis transmembrane conductance regulator (CFTR) biomarkers recommended to make a diagnosis in individuals with inconclusive sweat test and genetics and a clinical suspicion for cystic fibrosis (CF) or CFTR-related disorder (CFTR-RD).

Methods: NPD and ICM were measured according to standard operating procedures of the European Cystic Fibrosis Society Diagnostic Network Working Group.

Results: We assessed 219 individuals by NPD or ICM who had been referred to our laboratory due to clinical symptoms suggestive of CF, but inconclusive sweat test and genetics (median age: 16.

Functional analysis of the p.[Arg74Trp;Val201Met;Asp1270Asn]/p.Phe508del CFTR mutation genotype in human native colon.

Background: The impact of complex alleles on CFTR processing and function has yet not been investigated in native human tissue.

Methods: Intestinal current measurements (ICM) followed by CFTR immunoblot were performed on rectal biopsies taken from two siblings who are compound heterozygous for the CFTR mutations p.Phe508del and the complex allele p.

Key role of an ADP - ribose - dependent transcriptional regulator of NAD metabolism for fitness and virulence of Pseudomonas aeruginosa.

NAD is an essential co-factor of redox reactions and metabolic conversions of NAD-dependent enzymes. NAD biosynthesis in the opportunistic pathogen Pseudomonas aeruginosa has yet not been experimentally explored. The in silico search for orthologs in the P.

Expression of p53, Ki67, EcPV2- and EcPV3 DNA, and viral genes in relation to metastasis and outcome in equine penile and preputial squamous cell carcinoma.

Reasons For Performing Study: Equine penile and preputial squamous cell carcinoma (SCC) is a potentially lethal disease of which little is known regarding the relationship between tumour characteristics and prognosis.

Objectives: To assess the relationship between tumour differentiation grade (tumour subtype), presence of papillomaviruses, expression of viral genes (E2, E6, L1), nuclear proteins p53 and Ki67 and metastasis in equine penile and preputial SCC and to assess the relationship of tumour subtype, presence of papillomavirus type 2, p53 and Ki67 with survival.

Study Design: Retrospective case-control study using archived material.

The CF-modifying gene EHF promotes p.Phe508del-CFTR residual function by altering protein glycosylation and trafficking in epithelial cells.

The three-base-pair deletion c.1521_1523delCTT (p.Phe508del, F508del) in the cystic fibrosis transmembrane conductance regulator (CFTR) is the most frequent disease-causing lesion in cystic fibrosis (CF).

The effects of three-month oral supplementation with a nutraceutical and exercise on the locomotor pattern of aged horses.

Reasons For Performing Study: Multiple in vitro studies assessing articular tissues have indicated that glucosamine and chondroitin sulphate may possess anti-inflammatory effects, but little is known of their clinical effects in vivo. Many old horses have stiff joints, which is likely to be attributable to inflammation and therapy with these nutraceutical compounds could improve joint function.

Objectives: To assess the clinical effects of a mixed supplement on the improvement of stiff gait in aged horses.

Expression of cyclo-oxygenases-1 and -2, and microsomal prostaglandin E synthase-1 in penile and preputial papillomas and squamous cell carcinomas in the horse.

Reasons For Performing Study: Penile and preputial papilloma and squamous cell carcinoma (SCC) are commonly diagnosed in horses. Papillomas have the potential to progress to potentially lethal SCC. Knowledge of pathogenetic mechanisms may help in prevention and definition of treatment targets.

Systemic or intrasynovial medication as singular or as combination treatment in horses with (peri-)synovial pain.

Records of lame horses (2004-2007) with (peri-)synovial pain of the metacarpophalangeal joint (MCPJ, n = 53), digital flexor tendon sheath (DFTS, n = 20), and distal interphalangeal joint (DIPJ, n = 31) were evaluated (n = 104). Cases were grouped according to the following treatments: (1) systemic administration of NSAIDs (n = 40); (2) intrasynovial administration of glucocorticosteroids (n = 30); (3) combination treatment (n = 34). Outcome was defined as successful when the owner reported that the horse had returned to its previous level of performance within a period of up to six months after treatment.

Immunochemical analysis of mutant CFTR in lung explants.

Background/aims: Knowledge about the abundance and distribution of CFTR protein glycoforms in native lung tissue is scarce. For upcoming studies with correctors and potentiators for CFTR it is important to get more information about mutant CFTR protein biochemistry. Target for novel treatment is the most afflicted organ in cystic fibrosis (CF), the lung.

The effect of previous conditioning exercise on diaphyseal and metaphyseal bone to imposition and withdrawal of training in young Thoroughbred horses.

This study recorded the response to training of the diaphysis of the proximal phalangeal bone and the third metacarpal bone (Mc3) and the Mc3 proximal metaphysis. Nineteen 2- and 3-year old horses in training were exposed either to spontaneous exercise at pasture (PASTEX group) or additional imposed exercise (CONDEX group) from a very young age. Quantitative computed tomography scans were analysed for bone mineral content, size, bone mineral density, periosteal and endosteal circumference, cortical thickness and an estimate of bone strength.

Mild exercise early in life produces changes in bone size and strength but not density in proximal phalangeal, third metacarpal and third carpal bones of foals.

Exercise or lack of it in early life affects chondro-osseous development. Two groups of horses were used to investigate the effects of age and exercise regimen on bone parameters of diaphyseal, metaphyseal, epiphyseal and cuboidal bones of the distal limb of Thoroughbreds. One group had exercised only spontaneously from an early age at pasture (PASTEX group), while the other group of horses were exposed to a 30% greater workload through additional defined exercise (CONDEX).

Penile and preputial tumours in the horse: literature review and proposal of a standardised approach.

Penile and preputial tumours are not uncommon in the horse, but can cause discomfort and lead to serious complications. Several types of tumour of the male external genitalia have been described. The most common type is the squamous cell carcinoma (SCC), which is found mainly in older horses.

Amino acid profile during exercise and training in Standardbreds.

The objective of this study is to assess the influence of acute exercise, training and intensified training on the plasma amino acid profile. In a 32-week longitudinal study using 10 Standardbred horses, training was divided into four phases, including a phase of intensified training for five horses. At the end of each phase, a standardized exercise test, SET, was performed.

Functional analysis of F508del CFTR in native human colon.

The major cystic fibrosis mutation F508del has been classified by experiments in animal and cell culture models as a temperature-sensitive mutant defective in protein folding, processing and trafficking, but literature data on F508del CFTR maturation and function in human tissue are inconsistent. In the present study the molecular pathology of F508del CFTR was characterized in freshly excised rectal mucosa by bioelectric measurement of the basic defect and CFTR protein analysis by metabolic labelling or immunoblot. The majority of investigated F508del homozygous subjects expressed low amounts of complex-glycosylated mature F508del CFTR and low residual F508del CFTR-mediated chloride secretory activity in the rectal mucosa.

The propagation of induced tendon lesions in the equine superficial digital flexor tendon: an ex vivo study.

Reasons For Performing Study: Clinical tendon lesions usually enlarge during the first days to weeks after sustaining the injury due to enzymatic and biomechanical influences. Limiting this enlargement would positively influence prognosis related to lesion size.

Objectives: To investigate the effect of cyclic loading on the propagation of enzymatically and physically induced tendon lesions and to assess the effect of immobilisation thereon in an ex vivo model.

The effect of platelet-rich plasma on the neovascularization of surgically created equine superficial digital flexor tendon lesions.

The repair process of tendon injuries, which are common in both human and equine athletes, is slow and the quality of the repair tissue is often inferior to the original tendon tissue, which frequently leads to re-injury. The relatively poor vascularization of tendons is considered to be one of the reasons for their limited healing potential. Recently, platelet-rich plasma (PRP), an autologous concentrate of platelets, rich in growth factors, has been shown to enhance the repair process of injured tendons.

Computerised analysis of standardised ultrasonographic images to monitor the repair of surgically created core lesions in equine superficial digital flexor tendons following treatment with intratendinous platelet rich plasma or placebo.

The effectiveness of new therapies to treat tendon injuries is difficult to determine and is often based on semi-quantitative methods, such as grey level analysis of ultrasonographic images or subjective pain scores. The alternatives are costly and long-lasting end-stage studies using experimental animals. In this study, a method of ultrasonographic tissue characterisation (UTC), using mathematical analysis of contiguous transverse ultrasonographic images, was used for intra-vital monitoring of the healing trajectory of standardised tendon lesions treated with platelet rich plasma (PRP) or placebo.

Effects of platelet-rich plasma on the quality of repair of mechanically induced core lesions in equine superficial digital flexor tendons: A placebo-controlled experimental study.

Tendon injuries are notorious for their slow and functionally inferior healing. Intratendinous application of platelet-rich plasma (PRP) has been reported to stimulate the repair process of tendon injuries, but there is little conclusive evidence for its effectiveness. A placebo-controlled experimental trial was performed to test the hypothesis that a single intratendinous PRP treatment enhances the quality of tendon repair, as evidenced by improved biochemical, biomechanical, and histological tissue properties.

Overtrained horses alter their resting pulsatile growth hormone secretion.

The influence of intensified and reduced training on nocturnal growth hormone (GH) secretion and elimination dynamics was studied in young (1.5 yr) Standardbred geldings to detect potential markers indicative for early overtraining. Ten horses trained on a treadmill for 32 wk in age-, breed-, and gender-matched fixed pairs.

Multiple roles of Pseudomonas aeruginosa TBCF10839 PilY1 in motility, transport and infection.

Polymorphonuclear neutrophils are the most important mammalian host defence cells against infections with Pseudomonas aeruginosa. Screening of a signature tagged mutagenesis library of the non-piliated P. aeruginosa strain TBCF10839 uncovered that transposon inactivation of its pilY1 gene rendered the bacterium more resistant against killing by neutrophils than the wild type and any other of the more than 3000 tested mutants.

(Over)training effects on quantitative electromyography and muscle enzyme activities in standardbred horses.

Too intensive training may lead to overreaching or overtraining. To study whether quantitative needle electromyography (QEMG) is more sensitive to detect training (mal)adaptation than muscle enzyme activities, 12 standardbred geldings trained for 32 wk in age-, breed-, and sex-matched fixed pairs. After a habituation and normal training (NT) phase (phases 1 and 2, 4 and 18 wk, respectively), with increasing intensity and duration and frequency of training sessions, an intensified training (IT) group (phase 3, 6 wk) and a control group (which continued training as in the last week of phase 2) were formed.

Expression levels of FAS are regulated through an evolutionary conserved element in intron 2, which modulates cystic fibrosis disease severity.

We have analyzed frequent naturally occurring variants in the autogene FAS in two independent cystic fibrosis (CF) patient populations. Analysis of FAS expression levels from intestinal epithelial biopsies from 16 unrelated F508del-CFTR homozygotes showed a correlation between FAS intron 2 SNP rs7901656 and signals for Affymetrix GeneChip U133 Plus 2.0 probeset 204781_s_at consistent with a dominant model (P=0.

Evaluation of a new strategy to modulate skeletal development in Thoroughbred performance horses by imposing track-based exercise during growth.

Reason For Performing Study: No data exist on the intensity of exercise required or on possible harmful effects of increasing exercise in foals over the natural level when free at pasture.

Objectives: To investigate whether an increase in workload over free pasture exercise in the period from directly after birth to the start of training is tolerated by Thoroughbred (TB) foals without increasing injury rate or producing other undesired side effects.

Methods: Thirty-three TB foals were allocated to one of 2 exercise groups directly after birth.