Update on iodine-refractory differentiated thyroid carcinoma.

Radioiodine therapy represents a fundamental pillar in the routine adjuvant therapy of patients with high-risk differentiated thyroid carcinoma. However, a non-negligible percentage of these patients will develop iodine refractoriness, showing a worse prognosis, as well a lower survival, which demonstrates a clear need to explore different therapeutic approaches. Iodine refractory patient treatment continues to be a challenge, currently having different novel therapeutic options that should be known by the different specialties related to differentiated thyroid carcinoma (DTC). The aim of this work is to review iodine refractory thyroid carcinoma treatment, focusing especially on the definition of iodine refractoriness, highlighting its importance due to its high mortality, and introducing the different therapeutic options available for these patients.

Role of systemic inflammatory factors in gastroenteropancreatic neuroendocrine tumors (GEP-NETs) treated with peptide receptor radionuclide therapy (PRRT): From biology to theragnosis.

AIM: Systemic inflammatory factors have been validated as indicators of ongoing systemic inflammation that could be predictive markers of poor prognosis for oncological outcomes. However, the prognostic impact of systemic inflammation markers is unknown in patients with gastroenteropancreatic neuroendocrine tumors (GEP-NETs) treated with peptide receptor radionuclide therapy (PRRT). METHODS: We conducted an observational, retrospective, multicentric study of 40 patients with GEP or unknown origin NETs treated with PRRT between 2016 and 2020. The systemic inflammatory markers were calculated as follows: neutrophil to lymphocyte ratio (NLR)=neutrophil count/lymphocyte count, monocyte to lymphocyte ratio (MLR)=monocyte count/lymphocyte count, platelet to lymphocyte ratio (PLR)=platelet count/lymphocyte count, albumin to lymphocyte ratio (ALR)=albumin levels/lymphocyte count and derived Neutrophil to Lymphocyte ratio (dNLR)=neutrophil count/(leucocytes count - neutrophils count). Baseline analysis and after the second dose were used for the calculation of different ratios. RESULTS: The median age was 63 years (range 41-85), 55% were male. The baseline cut-off values for NLR were 2.61, for MLR 0.31, for PLR 110.14, for ALR 2.39 and for dNLR 1.71. The cut-off values after the 2° dose were, for NLR 2.3, for MLR 0.3, for PLR 131.61, ALR 4.16, and dNLR 1.48. Median progression-free survival (PFS) was 21.7 months (95% CI 10.7-32.8 months) and overall survival (OS) was 32.1 months (95% CI 19.6-44.7 months), PFS was shorter in patients with elevated NLR (p=0.001), ALR (0.03), and dNLR (p=0.001) in baseline analysis. DCR was 81% and ORR 18%. CONCLUSIONS: In GEP or unknown origin NETs treated with PRRT, we have identified the predictive and prognostic impact of baseline systemic inflammatory factors.

Biomechanical behavior of an alveolar graft under maxillary therapies.

Cleft lip and palate is a congenital defect that affects the oral cavity. Depending on its severity, alveolar graft surgery and maxillary orthopedic therapies must be carried out as a part of the treatment. It is widely accepted that the therapies should be performed before grafting. Nevertheless, some authors have suggested that mechanical stimuli such as those from the maxillary therapies could improve the success rate of the graft. The aim of this study is to computationally determine the effect of maxillary therapies loads on the biomechanical response of an alveolar graft with different degrees of ossification. We also explore how the transverse width of the cleft affects the graft behavior and compare results with a non-cleft skull. Results suggest that stresses increase within the graft as it ossifies and are greater if maxillary expansion therapy is applied. This has consequences in the bone remodeling processes that are necessary for the graft osseointegration. Maxillary orthopedic therapies after graft surgery could be considered as a part of the treatment since they seem to act as a positive extra stimulus that can benefit the graft.

Leptospirosis in an asplenic patient -case report.

BACKGROUND: The presentation of clinical leptospirosis has been historically associated with animal workers, slaughterhouse workers and medical veterinarians. This association has shifted to be related to flooding events and outdoor activities; few cases are related to high-risk factors found in immunosuppressed patients. Scarcely a handful of cases have serological evidence of immune response against Leptospira serovar Bratislava representing serogroup Australis, a serovar associated with poor reproductive performance in swine and horses, and recently with cats. CASE PRESENTATION: Herein, we describe a rare clinical presentation of disseminated Leptospira infection in an immunosuppressed 65-year-old woman. She was admitted to the emergency room with fever, bacteraemia, bilateral uveitis and pulmonary involvement. The patient denied outdoor activities; she only had wide exposure to faeces and urine from cats living in her home. Her medical history included idiopathic thrombocytopenic purpura (ITP) diagnosed at the age of 18. She did not respond to medical treatment, and a splenectomy was performed. At age 60, she was diagnosed with Chronic Myeloid Leukemia (CML), and was treated with a tyrosine kinase inhibitor (TKI) -Imatinib. The patient voluntarily discontinued the treatment for the last 6 months. After extensive workup, no microorganisms were identified by the commonly used stains in microbiology. The diagnosis was performed through dark-field microscopy, microagglutination test (MAT), Leptospira genus-specific PCR, the IS1500 PCR for identification of pathogenic species, and 16S based sequencing for the genus identification. CONCLUSION: Immunosuppressed patients may acquire uncommon infections from ubiquitous microorganisms. In this case, serology evidence of exposure to Leptospira serovar Bratislava by MAT and the presence of the Leptospira genus were identified. It should be on mind for the diagnosis in otherwise healthy patients, and thoroughly search on splenectomised patients exposed to animals. Additionally, this report highlights the usefulness of PCR for diagnosis of this potentially life-threatening illness.

Differential phenotype of immune cells in blood and milk following pegylated granulocyte colony-stimulating factor therapy during a chronic Staphylococcus aureus infection in lactating Holsteins.

Neutrophils are principal host innate immune cell responders to mastitis infections. Thus, therapies have been developed that target neutrophil expansion. This includes the neutrophil-stimulating cytokine granulocyte colony-stimulating factor (gCSF). Pegylated gCSF (PEG-gCSF; Imrestor, Elanco Animal Health, Greenfield, IN) has been shown to reduce the natural incidence of mastitis in periparturient cows in commercial settings and reduce severity of disease against experimental mastitis challenge. Pegylated gCSF stimulates neutrophil expansion but also induces changes in monocyte and lymphocyte circulating numbers, surface protein expression changes, or both. We hypothesized that PEG-gCSF modulates surface expression of monocytes and neutrophils and facilitates their migration to the mammary gland. We challenged 8 mid-lactation Holsteins with approximately 150 cfu of Staphylococcus aureus (Newbould 305) in a single quarter via intramammary infusion. All animals developed chronic infections as assessed by bacteria counts and somatic cell counts (SCC). Ten to 16 wk postchallenge, 4 of the animals were treated with 2 subcutaneous injections of PEG-gCSF 7 d apart. Complete blood counts, SCC, bacterial counts, milk yield, feed intake, neutrophils extracellular trap analysis, and flow cytometric analyses of milk and blood samples were performed at indicated time points for 14 d after the first PEG-gCSF injection. The PEG-gCSF-treated cows had significantly increased numbers of blood neutrophils and lymphocytes compared with control cows. Flow cytometric analyses revealed increased surface expression of myeloperoxidase (MPO) on neutrophils and macrophages in milk but not in blood of treated cows. Neutrophils isolated from blood of PEG-gCSF-treated cows had decreased surface expression of CD62L (L-selectin) in blood, consistent with cell activation. Surprisingly, CD62L cell surface expression was increased on neutrophils and macrophages sourced from milk from treated animals compared with cells isolated from controls. The PEG-gCSF-treated cows did not clear the S. aureus infection, nor did they significantly differ in SCC from controls. These findings provide evidence that PEG-gCSF therapy modifies cell surface expression of neutrophils and monocytes. However, although surface MPO+ cells accumulate in the mammary gland, the lack of bacterial control from these milk-derived cells suggests an incomplete role for PEG-gCSF treatment against chronic S. aureus infection and possibly chronic mammary infections in general.

Treponema pallidum ssp. pallidum identification by real-time PCR targetting the polA gene in paraffin-embedded samples positive by immunohistochemistry.

Syphilis is a systemic and sexually transmitted infection caused by Treponema pallidum ssp. pallidum. This spirochete causes different clinical and subclinical stages depending on the duration of infection and immune status of the host. Several tests have been developed for diagnosis, and are classified into direct and indirect methods. The first one includes dark field microscopy, direct fluorescent antibody test in fluids or tissue, and molecular biology techniques. In the indirect method (serologic), the routine tests are used, and are divided in two categories: non-treponemal and treponemal ones. The objective of this work was to identify T. pallidum ssp. pallidum in paraffin-embedded skin biopsies positive by immunohistochemistry, using conventional polymerase chain reaction (PCR) and quantitative real time PCR (qPCR). We included a sample of 17 paraffin-embedded biopsies. DNA was extracted and processed by conventional PCR and real-time PCR with a TaqMan® probe to identify the polA gene. Using PCR, 11 tested positive (64.7%) and 6 (35.3%) were negative. With qPCR and TaqMan® probe, 100% of samples tested positive. The minimum number of spirochetes detected in each sample was 2. With this work, we can conclude that qPCR is a fast and very accurate method for diagnosis of syphilis in tissue specimens.

Defective sarcoplasmic reticulum-mitochondria calcium exchange in aged mouse myocardium.

Mitochondrial alterations are critically involved in increased vulnerability to disease during aging. We investigated the contribution of mitochondria-sarcoplasmic reticulum (SR) communication in cardiomyocyte functional alterations during aging. Heart function (echocardiography) and ATP/phosphocreatine (NMR spectroscopy) were preserved in hearts from old mice (>20 months) with respect to young mice (5-6 months). Mitochondrial membrane potential and resting O2 consumption were similar in mitochondria from young and old hearts. However, maximal ADP-stimulated O2 consumption was specifically reduced in interfibrillar mitochondria from aged hearts. Second generation proteomics disclosed an increased mitochondrial protein oxidation in advanced age. Because energy production and oxidative status are regulated by mitochondrial Ca2+, we investigated the effect of age on mitochondrial Ca2+ uptake. Although no age-dependent differences were found in Ca2+ uptake kinetics in isolated mitochondria, mitochondrial Ca2+ uptake secondary to SR Ca2+ release was significantly reduced in cardiomyocytes from old hearts, and this effect was associated with decreased NAD(P)H regeneration and increased mitochondrial ROS upon increased contractile activity. Immunofluorescence and proximity ligation assay identified the defective communication between mitochondrial voltage-dependent anion channel and SR ryanodine receptor (RyR) in cardiomyocytes from aged hearts associated with altered Ca2+ handling. Age-dependent alterations in SR Ca2+ transfer to mitochondria and in Ca2+ handling could be reproduced in cardiomyoctes from young hearts after interorganelle disruption with colchicine, at concentrations that had no effect in aged cardiomyocytes or isolated mitochondria. Thus, defective SR-mitochondria communication underlies inefficient interorganelle Ca2+ exchange that contributes to energy demand/supply mistmach and oxidative stress in the aged heart.

Polymorphisms in calpastatin and mu-calpain genes are associated with beef iron content.

The objective of this study was to assess the association of markers in the calpastatin and mu-calpain loci with iron in beef cattle muscle. The population consisted of 259 cross-bred steers from Beefmaster, Brangus, Bonsmara, Romosinuano, Hereford and Angus sires. Total iron and heme iron concentrations were measured. Markers in the calpastatin (referred to as CAST) and mu-calpain (referred to as CAPN4751) genes were used to assess their association with iron levels. The mean and standard error for iron and heme iron content in the population was 35.6 ± 1.3 µg and 27.1 ± 1.4 µg respectively. Significant associations (P < 0.01) of markers were observed for both iron and heme iron content. For CAST, animals with the CC genotype had higher levels of iron and heme iron in longissimus dorsi muscle. For CAPN4751, individuals with the TT genotype had higher concentrations of iron and heme iron than did animals with the CC and CT genotypes. Genotypes known to be associated with tougher meat were associated with higher levels of iron concentration.

Análise biomecânica ex vivo de um modelo de haste intramedular de polipropileno para osteossíntese em úmeros de bezerros / Ex vivo biomechanics assay of a model of polypropylene intramedullary nail for calf humeral osteosynthesis

Foram utilizados 12 pares de úmeros obtidos de bezerros machos da raça Holandesa, com idades entre 15 e 30 dias. Os úmeros esquerdos foram mantidos íntegros, e os direitos foram fraturados de forma oblíqua em sua diáfise, na transição entre os terços médio e proximal. A redução da fratura foi feita pela aplicação intramedular de haste de polipropileno, bloqueada por dois parafusos corticais de aço inoxidável, dispostos transversalmente em cada fragmento ósseo. Seis pares de ossos foram submetidos ao teste de compressão, e seis ao teste de flexão, utilizando-se uma máquina universal de ensaios. Nos testes de compressão, as cargas médias de ruptura foram 738,3N e 473,3N, e nos testes de flexão 322,4N e 117,9N, para os ossos íntegros e fraturados, respectivamente. Comparando-se o grupo de ossos fraturados com o grupo de ossos íntegros, verificou-se que o sistema proposto foi capaz de resistir a 66,4 por cento das cargas médias quando submetido à compressão, e a 36,6 por cento quando submetido à flexão. Úmeros fraturados e tratados com haste intramedular de polipropileno apresentaram resistência limitada se comparados aos ossos íntegros.

A total of 12 pairs of humeri from male calves 15 to 30 days old were used. The left humeri remained unchanged and the right ones were fractured in the diaphysis between proximal and middle thirds. The fracture was fixed with a polypropylene intramedullary nail interlocked with two steel bone screws crossed along each segment. Six pairs of bones were subjected to a compression test and the others to a flexural test using a universal testing machine. In the compression tests, the mean rupture loads were 738,3N and 473,3N, and in the flexural tests they were 322,4N and 117,9N for the intact and fractured bones respectively. Fractured bones fixed with the proposed model were able to resist 66.4 percent of the load during compression and 36.6 percent during bending when in comparison to intact bones. Fractured humeri treated with polypropylene intramedullary nail showed limited resistance compared to intact ones.

Bone remodelling analysis of a bovine femur for a veterinary implant design.

The response of bovine bone to the presence of an implant is analysed with the aim of simulating bone remodelling in a developing model of a polymeric intramedullary interlocking nail for veterinary use. A 3-D finite element model of the femur diaphysis is built based on computed tomography images and using a CAD-based modelling pipeline. The bone remodelling process after the surgery is analysed and compared with the healthy bone. The remodelling law assumes that bone adapts to the mechanical environment. For the analyses a consistent set of loads is determined for the bovine walk cycle. The remodelling results reproduce the morphologic features of bone and provide evidence of the difference on the bone behaviour when comparing metallic and polymeric nails. Our findings indicate that an intramedullary polymeric nail has the advantage over the metallic one of improving long-term bone healing and possibly avoiding the need of the implant removal.

Predicting gene promoter methylation in non-small-cell lung cancer by evaluating sputum and serum.

The use of 5-methylcytosine demethylating agents in conjunction with inhibitors of histone deacetylation may offer a new therapeutic strategy for lung cancer. Monitoring the efficacy of gene demethylating treatment directly within the tumour may be difficult due to tumour location. This study determined the positive and negative predictive values of sputum and serum for detecting gene methylation in primary lung cancer. A panel of eight genes was evaluated by comparing methylation detected in the primary tumour biopsy to serum and sputum obtained from 72 patients with Stage III lung cancer. The prevalence for methylation of the eight genes in sputum (21-43%) approximated to that seen in tumours, but was 0.7-4.3-fold greater than detected in serum. Sputum was superior to serum in classifying the methylation status of genes in the tumour biopsy. The positive predictive value of the top four genes (p16, DAPK, PAX5 beta, and GATA5) was 44-72% with a negative predictive value for these genes > or =70%. The highest specificity was seen for the p16 gene, and this was associated with a odds ratio of six for methylation in the tumour when this gene was methylated in sputum. In contrast, for serum, the individual sensitivity for all genes was 6-27%. Evaluating the combined effect of methylation of at least one of the four most significant genes in sputum increased the positive predictive value to 86%. These studies demonstrate that sputum can be used effectively as a surrogate for tumour tissue to predict the methylation status of advanced lung cancer where biopsy is not feasible.

Association of markers in the bovine CAPN1 gene with meat tenderness in large crossbred populations that sample influential industry sires.

Two previously identified single-nucleotide polymorphism markers located within the micromolar calcium-activated neutral protease gene (CAPN1) were evaluated for their association with variation in meat tenderness using one commercial sample of Simmental x Angus crossbred calves and one multibreed, crossbred research herd. The commercial sample included 362 animals sired by 23 registered Simmental bulls bred to unregistered Angus cows and represented current industry animals in which to test the predictive merit of the markers. The second sample was a research herd including 564 steers from the Germplasm Evaluation Cycle VII population at the U.S. Meat Animal Research Center, produced with semen from popular sires of the seven Bos taurus beef breeds with the most registrations in the United States (Angus, Charolais, Gelbvieh, Hereford, Limousin, Red Angus, and Simmental) on Angus, Hereford, and MARC III cows. These animals form a relatively outbred population that constituted a stringent test of the predictive merit of the genetic markers, although small groups were half-sibs. Warner-Bratzler shear force measurements were used to determine tenderness phenotypes for all animals. The populations were genotyped for two markers that predict variation at amino acid positions 316 and 530 of the mu-calpain polypeptide, produced by the CAPN1 gene. Minor allele frequencies for markers 316 and 530 in the commercial sample were 0.17 and 0.37, respectively, and in the Cycle VII animals, were 0.20 and 0.28, respectively. Both markers showed association with shear force in the commercial sample (P = 0.04) and the Cycle VII population (P = 0.02), supporting the hypothesis that they represent potential markers to aid selection for improved meat tenderness in commercial populations of beef cattle in the United States.

Tyrosol and hydroxytyrosol are absorbed from moderate and sustained doses of virgin olive oil in humans.

OBJECTIVE: To investigate the absorption of tyrosol and hydroxytyrosol from moderate and sustained doses of virgin olive oil consumption. The study also aimed to investigate whether these phenolic compounds could be used as biomarkers of virgin olive oil intake. DESIGN AND INTERVENTIONS: Ingestion of a single dose of virgin olive oil (50 ml). Thereafter, for a week, participants followed their usual diet which included 25 ml/day of the same virgin olive oil as the source of raw fat. SETTING: Unitat de Recerca en Farmacologia. Institut Municipal d'Investigació Mèdica (IMIM). SUBJECTS: Seven healthy volunteers. RESULTS: An increase in 24 h urine of tyrosol and hydroxytyrosol, after both a single-dose ingestion (50 ml) and short-term consumption (one week, 25 ml/day) of virgin olive oil (P<0.05) was observed. Urinary recoveries for tyrosol were similar after a single dose and after sustained doses of virgin olive oil. Mean recovery values for hydroxytyrosol after sustained doses were 1.5-fold those obtained after a single 50 ml dose. CONCLUSIONS: Tyrosol and hydroxytyrosol are absorbed from realistic doses of virgin olive oil. With regard to the dose-effect relationship, 24 h urinary tyrosol seems to be a better biomarker of sustained and moderate doses of virgin olive oil consumption than hydroxytyrosol.

Bioavailability of tyrosol, an antioxidant phenolic compound present in wine and olive oil, in humans.

Tyrosol is a phenolic compound present in two of the traditional components of the Mediterranean diet: wine and virgin olive oil. The presence of tyrosol has been described in red and white wines. Tyrosol is also present in vermouth and beer. Tyrosol has been shown to be able to exert antioxidant activity in in vitro studies. Oxidation of low-density lipoprotein (LDL) appears to occur predominantly in arterial intima in microdomains sequestered from antioxidants of plasma. The antioxidant content of the LDL particle is critical for its protection. Thus, phenolics, which are able to bind LDL, could be effective in preventing lipid peroxidation and atherosclerotic processes. The ability of tyrosol to bind human LDL has been reported. We have demonstrated the bioavailability of tyrosol in humans from virgin olive oil in its natural form. Urinary tyrosol increased, reaching a peak at 0-4 h after virgin olive oil administration. Men and women showed a different pattern of urinary excretion of tyrosol. Moreover, tyrosol is absorbed in a dose-dependent manner after sustained and moderate doses of virgin olive oil. In summary, our results suggest that tyrosol from wine or virgin olive oil could exert beneficial effects on human health in vivo if its biological properties are confirmed in in vivo studies.

Capillary gas chromatography-mass spectrometry quantitative determination of hydroxytyrosol and tyrosol in human urine after olive oil intake.

Recent in vitro studies have demonstrated antioxidant properties of some virgin olive oil phenolic compounds. One of the prerequisites to extrapolate these data to an in vivo situation is the knowledge of their bioavailability in humans. In the present work we describe an analytical method which enables us to perform hydroxytyrosol and tyrosol quantitative determinations in human urine. This method was successfully used in bioavailability studies of both phenolic compounds after acute olive oil administration. Virgin olive oil was administered to healthy volunteers after a low phenolic diet. The dose administered of both phenolic compounds was estimated in reference to free forms of hydroxytyrosol and tyrosol present in virgin olive oil extracts before and after being submitted to hydrolytic conditions. These conditions mimic those occurring during digestion. Urine samples were collected before and after acute olive oil intake and analyzed by capillary gas chromatography-mass spectrometry. Hydroxytyrosol and tyrosol urinary recovery increased in response to olive oil administration, obtaining maximal values in the first 4 h. Our results further indicate that hydroxytyrosol and tyrosol are mainly excreted in conjugated form, since only 5.9 +/- 1.4% (hydroxytyrosol) and 13.8 +/- 5.4% (tyrosol) of the total amounts excreted in urine were in free form.

Changes in cyclin B localisation during pig oocyte in vitro maturation.

The localisation of cyclin B throughout in vitro maturation of pig oocytes was determined by indirect immunofluorescence using a monoclonal antibody specific for an epitope of the human cyclin B. Maturation of pig oocytes was induced by addition of Pergonal (2 UI/ml of FSH/LH) and beta-oestradiol to the medium where isolated ovarian follicles were cultured for up to 72 h. Immature gametes with an intact germinal vesicle were observed during the first 30 h of culture. Only 10% were competent to reinitiate meiosis and showed germinal vesicle breakdown (GVBD) after 36 h. However, after 48-72 h, 60% of the oocytes accomplished their maturation and showed metaphase chromosomes. Immature oocytes showed cyclin B immunofluorescent staining in the cytoplasm, whereas mature oocytes showed the immunofluorescent label concentrated in the nucleus. Metaphase chromosomes showed an intense immunofluorescence. The migration of cyclin B to the nucleus and its association with metaphase chromosomes in pig oocytes able to progress through meiosis resembled the subcellular localisation of cyclin B and the distribution of maturation promoting factor (MPF) in mitotic dividing cells.