Allele frequency of muscular genetic disorders in bull-catching (vaquejada) quarter horses.

Quarter horses (QH), a prominent athletic breed in Brazil, are affected by muscular genetic disorders such as myosin-heavy chain myopathy (MYHM), polysaccharide storage myopathy (PSSM1), hyperkalemic periodic paralysis (HyPP), and malignant hyperthermia (MH). Bull-catching (vaquejada), primarily involving QH, is a significant equestrian sport in Brazil. Since the allele frequencies (AF) of MYHM, PSSM1, HyPP, and MH in vaquejada QH remain unknown, this study evaluated the AF in 129 QH vaquejada athletes, specifically from the Brazilian Northeast. These variants were exclusively observed in heterozygosity. The MYHM exhibited the highest AF (0.04 ±0.01), followed by PSSM1 (0.01 ±0.01) and the HyPP variant (0.004 ±0.01), while the MH variant was not identified in this study. This study represents the first identification of these variants in vaquejada QH, emphasizing the need to implement measures to prevent the transmission of pathogenic alleles and reduce the occurrence of clinical cases of these genetic diseases.

Blood and sperm traces on human hair. A study on preservation and detection after 3-month outdoor exposure.

Hair can retain important biological traces for forensic investigations. Forensic scientists are used to looking for such traces on clothing and skin of victims, however, when decomposition kicks in and all that remains of the victims is the skeleton, hair may be the only tissue representing the surface of the body at the time of a crime on which biological traces of an aggressor may have been left and still be detectable. Given the lack of research on this topic, this pilot study aims to assess the capacity of hair to retain semen and blood in hair, and the possibility to detect these fluids with well-known techniques and to obtain a useful genetic profile even when exposed to environmental conditions (Open Natural Environment (woods), Open Man Made Environment (urban)) for three months. Results showed that both traces were always visible and detectable with almost all techniques in the Control Environment, while in the two open environments some difficulties arose. However, biomolecular analysis was effective up to three months on both fluids in the Natural Environment and up to two months and one week respectively on blood and semen in the Man Made Environment. The Combur Test, OBTI, and Luminol were effective on blood up to three months in both environments while Sperm-HY-Liter and observation of cellular components were effective on semen up to at least 1 month and PSA testing was positive up to 1 week in both environments. The present work can be considered an encouraging starting point for the analysis of biological traces on hair in forensic contexts, regardless of the PMI, since blood and semen related to a crime may survive.

Blood thyroglobulin and TSH receptor mRNA detection by RT-PCR in the follow-up of differentiated thyroid cancer patients.

OBJECTIVE: During the last years several groups have used the technique of RT-PCR for the detection of circulating thyroid cells, through the amplification of thyroglobulin (Tg) and TSH receptor(TSH-R) mRNA; however the published results are controversial. In this study we investigated the utility for the detection of Tg and TSH-R mRNA by RT-PCR in patients with differentiated thyroid cancer (DTC) during treatment with levothyroxine. SUBJECTS AND METHODS: We investigated the expression of Tg and TSH-R mRNA by single and nested RT-PCR in the blood of 3 groups of subjects: (A) 34 patients with DTC and no evidence of disease, (B) 8 patients with DTC and evidence of local or distant metastasis and (C) 13 normal subjects. Expression levels of Tg mRNA were also analysed by comparative semi-quantitative RT-PCR. RESULTS: Tg and TSH-R mRNA signals were detected in all subjects (patients with DTC with and without evidence of disease and in normal subjects) by single or nested RT-PCR. By semi-quantitative RT-PCR and densitometric analysis of PCR products, mean levels of circulating Tg mRNA of the 3 groups were: Group A 0.182+/-0.107, Group B 0.329+/-0.298 and Group C 0.305+/-0.217. CONCLUSIONS: Single or nested RT-PCR for Tg and TSH-R mRNA is not a suitable tool in the follow-up of patients with DTC. Lower levels of Tg mRNA in patients with DTC without evidence of disease, although not significant, may indicate that small numbers of thyroid cells may be normally present in the circulation or may represent an ectopic transcription of messengers from blood cells.

[Regional cerebral blood flow changes in Parkinson's disease: correlation with disease duration]. / Cambios de la perfusión cerebral en la enfermedad de Parkinson: relación con la duración de los síntomas.

INTRODUCTION: Changes in regional cerebral blood flow (rCBF) have been reported in idiopathic Parkinson's disease (PD). Nonetheless, their typical pattern still remains controversial regarding some features, such as basal ganglia involvement and the main cortical regions affected. Functional neuroimaging makes it possible to identify the brain dysfunctions of the neural circuits underlying the disease. Voxel-based analysis methods make it possible to increase the reliability of the results. OBJECTIVE: To assess the rCBF changes in patients with PD and their relation with disease duration. MATERIALS AND METHODS: Thirty PD adult patients without dementia underwent evaluation with (99m)Tc-ECD SPECT. SPM5 was used for statistical comparison with 25 normal controls of similar ages. The disease course duration in years was added as a covariate. Additionally, patients with a 6-year evolution or less and those with more than 6 years were compared separately with normal controls. RESULTS: Significant hypoperfusion was detected in bilateral premotor and posterior parietal cortex and increase of perfusion was present in the cerebellum. These changes correlated with the years of evolution of the illness. Patients with longer evolution also presented thalamic, subthalamic and basal ganglia hypoperfusion. CONCLUSIONS: We describe rCBF changes in PD in neural circuits related with control of movements. These changes are more manifest in patients with a longer duration of the disease.

Cutaneous lymphoscintigraphy in malignant melanoma.

Melanoma commonly metastasizes to the regional lymph nodes. With early melanomas the accepted therapy is local wide excision and regional lymphadenectomy for nodes in the lymph drainage path. For some melanoma sites the lymph node groups that should be removed are obvious; however, for other sites the decision as to which nodes are at risk may present a difficult problem, since drainage from these sites may vary unpredictably from classical anatomic descriptions. We reviewed recent reports from four institutions using four radiocolloids (colloidal 198gold, 99mtechnetium antimony sulfur colloid (99mTcSbSC), 99mtechnetium sulfur colloid (99mTcSC), and micro 99mtechnetium sulfur colloid (micro 99mTcSC)) to determine lymph shed from melanomas. Correlation with surgical findings was excellent. In addition, drainage to second and third nodal groups, some of which were not clinically predictable, was frequently observed. While all four agents gave good scans, 99mTcSC and micro 99mTcSC are probably the best radiopharmaceuticals.

Detection of recurrent malignant melanoma with 99mTc-MIBI scintigraphy.

Initial reports suggest that 99mTc-methoxyisobutylisonitrile (MIBI) scanning may be of clinical value in staging patients with malignant melanoma. We carried out a study to evaluate the potential of this technique in the detection of recurrent disease. Whole-body 99mTC-MIBI scans were performed in 81 patients with a history of a surgically excised MM: 28 with known recurrent lesions and 53 during follow-up without evidence of disease. Images started 10 min post-injection, using a dose of 740 MBq. Diagnoses were confirmed by cytological/histological examination or at least one conventional imaging modality. Blinded interpretations of the MIBI scans were performed. Whole-body MIBI scanning correctly detected 68 (92%) of 74 metastatic lesions in the following sites: regional lymph nodes (n=23), non-regional lymph nodes (n=10), skin (n=16), brain/cerebellum (n=6), lung (n=8), bone (n=4) and breast (n=1). The technique failed to detect three subcutaneous regressive lesions (< 1 cm), one liver metastasis, one spleen metastasis and a case of multiple small lesions of the duodenal mucous membrane. In 14 patients the procedure detected previously unknown metastatic lesions. These results suggest that 99mTc-MIBI scanning is an effective imaging modality for whole-body screening of metastatic disease in malignant melanoma patients with the potential to influence treatment planning.

99Tc(m)-MIBI scanning of the thyroid gland in patients with markedly decreased pertechnetate uptake.

Diffusely reduced 99Tc(m)-pertechnetate uptake is a relatively infrequent but annoying finding that impairs evaluation of the thyroid gland. We studied 32 female patients aged 19-85 years with markedly reduced pertechnetate uptake. The following causes of reduced pertechnetate uptake were recognized: treatment with iodinated pharmaceuticals (n = 15), suppression therapy with T4 (n = 11), subacute thyroiditis (n = 5) and massive tumour replacement (n = 1). 99Tc(m)-MIBI thyroid scintigraphy was performed within 24 h of the pertechnetate study. The results were correlated with neck ultrasound, serum TSH (n = 25) and surgical findings in patients who had been operated on. The technique identified the following conditions: normally sized thyroids (n = 4), diffuse goitres (n = 8), multinodular goitres (n = 17) and solitary thyroid nodules (n = 3). Moreover, substernal goitres were identified in nine patients. This condition was confirmed at surgery in seven patients. Ultrasonography was concordant in 29 of 32 patients in terms of thyroid size and structure, but failed to demonstrate substernal thyroid tissue. Our results suggest that 99Tc(m)-MIBI scintigraphy may contribute to the diagnosis of thyroid pathology and treatment planning in patients with diffusely decreased 99Tc(m)-pertechnetate uptake.

Italian population data on thirteen short tandem repeat loci: HUMTH01, D21S11, D18S51, HUMVWFA31, HUMFIBRA, D8S1179, HUMTPOX, HUMCSF1PO, D16S539, D7S820, D13S317, D5S818, D3S1358.

A population study on thirteen short tandem repeat (STR) loci was performed on 223 unrelated Italian Caucasians. The DNA was amplified by PCR. Separation and detection of the amplified STR fragments was carried out by use of 377 automated system (Applied Biosystems Division/Perkin Elmer). All loci meet Hardy-Weinberg expectations, and the data show only five departures out of seventy-eight pairwise locus tests which is close to expectations of 5% (5/78 = 6.4%). When correcting for multiple tests, there is little evidence for departures from expectations between loci. The combined Power of Exclusion for the thirteen STR loci is 0.99999270. The results demonstrate that these loci will be very useful for human identification in forensic cases in Italy.

Changes in the cerebral blood flow in postlingual cochlear implant users.

Five postlingually deaf patients (age range 28-58 years) with multichannel cochlear implants were examined with single photon emission tomography (SPECT) (triple-head rotating gamma camera). Changes in the regional cerebral blood flow (rCBF) after intravenous administration of technetium-99m ethyl cysteinate dimer (Tc-99m ECD) were assessed through a stimulation paradigm, consisting of: i) click stimuli (75 dB SPL) in the ear that was to be implanted, 2 weeks before surgery; ii) stimulation with the same click, one month after initial fitting; iii) stimulation with hearing sequential Spanish sentences one month after initial fitting. The results showed a significant increase in the rCBF in the primary left auditory area and in the right auditory cortex, in conditions ii) and iii). The rCBF also showed a significant asymmetrical increase in the frontal lobes when the patient was hearing sequential sentences (condition iii)) with asymmetrical distribution among patients. These results are discussed, principally the correlation between speech discrimination scores and the rCBF distribution in the frontal and temporal lobes.

Preoperative evaluation of sentinel lymph node status in melanoma by means of (99m)Tc-MIBI and lymphoscintigraphy: a case report.

Previous reports have shown that (99m)Tc-sestamibi (MIBI) could detect clinically occult metastatic melanoma lesions. This article reports on a patient with invasive melanoma of the right heel in whom the sentinel node status was preoperatively evaluated with this tracer. Although regional lymph nodes were clinically negative, (99m)Tc-MIBI scintigraphy showed focal increased tracer uptake in the right groin that corresponded to the location of 2 sentinel nodes visualized by lymphoscintigraphy with (99m)Tc-colloidal rhenium sulfide performed the same day. A gamma-probe was used intraoperatively to guide the excision of the sentinel nodes that were further classified as metastatic by histopathology. This double-technique approach is technically feasible and has the potential of selecting a group of patients who might benefit from a selective complete lymphadenectomy.

Evaluation of ocular tumors with technetium-99m-MIBI: planar pinhole technique or SPECT?

OBJECTIVE: This study compares 2 imaging protocols, planar pinhole technique (PPHT) and SPECT, for evaluating ocular masses with 99mTc-MIBI. METHODS: Sixteen patients with ocular lesions were studied. Planar images were acquired 10 min after the injection of 740 MBq 99mTc-MIBI with an LFOV camera fitted with a pinhole collimator (5.0 mm). A SPECT study was performed immediately after the planar study, using a 360 degrees orbit, 64 steps, 20 s/stop, a 128 x 128 matrix, and a low-energy high-resolution (LEHR) collimator. Twelve lesions (9.5-18.0 mm) proved to be malignant: 8 primary tumors (ocular melanoma); 3 local relapses of different tumors of the conjunctiva; and 1 ocular metastasis from breast cancer. The remaining 4 lesions (10.0-16.0 mm) were benign: 1 inflammatory lesion; 1 benign intraocular calcification; and 2 naevi. RESULTS: SPECT images showed 11 of 12 malignant lesions (91.6%), whereas the planar technique demonstrated only 4 of the 12 lesions (33.3%). One false-positive result, the inflammatory lesion, was visualized by both techniques. The remaining benign lesions were not detected with either method. CONCLUSION: Technetium-99m-MIBI SPECT is a sensitive technique for detecting malignant ocular tumors. SPECT imaging is a better alternative to planar imaging for ocular tumors.

Italian population data on the polymarker system and on the five short tandem repeat loci CSF1PO, TPOX, TH01, F13B, and vWA.

A population study on five short tandem repeat (STR) loci and five sequence specific polymorphism loci was performed on unrelated Italian Caucasians. Separation and detection of the amplified STR fragments were carried out by high resolution vertical denaturing polyacrylamide gel electrophoresis (PAGE) and silver staining, respectively. The sequence specific loci were analyzed using the AmpliType PM Typing Kit (Perkin Elmer, Foster City, CA). All loci, except Gc (p = 0.031), meet Hardy-Wienberg expectations. In addition, there is no evidence for association of alleles between pairs of loci. The combined power of discrimination for the five STR loci is 0.9999862 and for the PM loci is 0.99503. The results suggest that these loci may be useful for human identification cases in Italy.

Thyroid imaging with Tc-99m MIBI in patients with solitary cold single nodules on pertechnetate imaging.

Thyroid imaging was performed in 30 patients with the standard pertechnetate technique, as well as with Tc-99m MIBI using a double-phase acquisition protocol. All patients had normal thyroid function confirmed by hormone measurements and cold solitary thyroid nodules, which were evaluated by pertechnetate scanning. Tc-99m MIBI scans were reported as showing cold (N = 14), warm (N = 7), or hot (N = 9) nodules. Nodule classification was made according to fine needle aspiration biopsy findings in 20 patients. The remaining 10 proceeded to surgery and had histopathologic confirmation of their lesions. Although all cold nodules with Tc-99m MIBI were cystic, six of the warm nodules were benign lesions. No histologically proven benign nodule was hot with Tc-99m MIBI. Of the hot nodules, seven were suspicious for follicular carcinoma with fine needle aspiration biopsy (N = 3), or had histologically proven papillary carcinoma (N = 4). Delayed images in five of seven of these lesions showed nodular retention of the radiopharmaceutical. In conclusion, double-phase Tc-99m MIBI scanning of the thyroid gland could be helpful in the preoperative assessment of patients with cold solitary thyroid nodules in order to evaluate the malignancy probability of these lesions.

Scintigraphic detection of invasive cutaneous malignant melanoma with Tc-99m MIBI.

Malignant melanoma is a relatively uncommon cancer of increasing incidence. The authors report on two cases of malignant melanoma lesions (primary and lymph node metastasis) imaged with Tc-99m MIBI. One patient revealed intense accumulation of the radiotracer in a primary skin lesion of the thorax, whereas the other patient showed increased Tc-99m MIBI uptake in a lymph node metastasis of the neck. Tc-99m MIBI imaging could be helpful in the evaluation of patients with suspicious skin lesions and in patients with known cutaneous malignant melanoma, in the investigation of lymph nodal invasion, local recurrence, and metastatic spread.

Scintigraphic evaluation of malignant melanoma lesions with Tc-99m tetrofosmin.

Two cases of malignant melanoma (primary and metastatic lesions) imaged with Tc-99m tetrofosmin are reported. One patient showed intense uptake in a primary skin lesion of the thorax, and the other patient had accumulation in skin, cerebellum, breast, and lymph node metastases. Like Tc-99m MIBI, Tc-99m tetrofosmin imaging may be of clinical relevance in the evaluation of suspicious skin lesions and in patients with known cutaneous malignant melanoma in the assessment of recurrent disease during follow-up evaluation.

[Examination of regional limph nodes in penile carcinoma by 99M Tc-MIBI scintigraphy]. / Exploración de ganglios regionales en carcinoma de pene mediante gammagrafía con 99mTc-MIBI.

99mTc-MIBI scintigraphy has proven utility as a clinically valuable technique for the diagnosis of various primary and recurrent malignant lesions. We report the case of a 65 year old man with penile squamous cell carcinoma and clinically questionable bilateral inguinal lymph nodes. 99mTc-MIBI scan shows increased focal uptake in both groins and in the right iliac region. The patient underwent penile amputation with bilateral lymphadenectomy. Histopathologic report revealed metastatic lymph nodes in all lymphatic basins seen with 99mTc-MIBI. We conclude that this technique has the potential of supplying clinically relevant information for the diagnosis of regional extension of patients with penile carcinoma.

[Cutaneo-visceral disseminated flat angioma, glaucoma, hypospadias and testicular ectopy]. / Angioma plano diseminado cutáneo visceral, glaucoma, hipospadias y ectopia testicular.

A case of associated disease in a child is described. It consisted of a disseminated capillary naevus with a vascular malformation in the gastrointestinal tract, bilateral congenital glaucoma, hypospadias, and bilateral ectopic testicles.

NIR luminescence for the inspection of thermal paper: a novel tool for fingermarks detection.

The near infrared (NIR) spectral region could offer advantages over the visible region in the detection of latent fingermarks due to the enhanced contrast between the background and the print. In this present paper, this spectral range of the electromagnetic spectrum was explored in order to study the possibility to make feasible the inspection of forensic evidences when the substrate is represented by the thermal paper, which is one of the most extensively used porous surfaces for printing receipts. We successfully applied the method to thermal paper treated with several kinds of amino acid sensitive reagents. We demonstrated that, even if the treatments could cause a dark stain over the latent fingermarks, by exploiting the luminescence of thermal paper in the NIR window we were able to create a contrast otherwise impossible in the visible region.

Decellularized omentum as novel biologic scaffold for reconstructive surgery and regenerative medicine.

Homologous tissues, such as adipose tissue, may be an interesting source of acellular scaffolds, maintaining a complex physiological three-dimensional (3D) structure, to be recellularized with autologous cells. The aim of the present work is to evaluate the possibility of obtaining homologous acellular scaffolds from decellularization of the omentum, which is known to have a complex vascular network. Adult rat and human omenta were treated with an adapted decellularization protocol involving mechanical rupture (freeze-thaw cycles), enzymatic digestion (trypsin, lipase, deoxyribonuclease, ribonuclease) and lipid extraction (2-propanol). Histological staining confirmed the effectiveness of decellularization, resulting in cell-free scaffolds with no residual cells in the matrix. The complex 3D networks of collagen (azan-Mallory), elastic fibers (Van Gieson), reticular fibers and glycosaminoglycans (PAS) were maintained, whereas Oil Red and Sudan stains showed the loss of lipids in the decellularized tissue. The vascular structures in the tissue were still visible, with preservation of collagen and elastic wall components and loss of endothelial (anti-CD31 and -CD34 immunohistochemistry) and smooth muscle (anti-alpha smooth muscle actin) cells. Fat-rich and well vascularized omental tissue may be decellularized to obtain complex 3D scaffolds preserving tissue architecture potentially suitable for recellularization. Further analyses are necessary to verify the possibility of recolonization of the scaffold by adipose-derived stem cells in vitro and then in vivo after re-implantation, as already known for homologus implants in regenerative processes.

A novel ß-glucosidase from Sporidiobolus pararoseus: characterization and application in winemaking.

UNLABELLED: For the first time, the production of an extracellular ß-glucosidase (Sp-ß-gl) by a Sporidiobolus pararoseus yeast strain is reported. The Sp-ß-gl activity was quantified, characterized, and assessed for its efficiency in releasing aroma-enhancing compounds in wines. The maximum enzymatic synthesis was after 72 h of growth in a complex media with 20 g/L of cellobiose. The optimal pH and temperature were 5.5 and at 50 °C, respectively. It showed a wide range of pH stability and exhibited quite high thermostability at low temperatures. In addition, this ß-glucosidase revealed tolerance to wine-associated inhibitory compounds (sugars and ethanol), showing suitable characteristics for all the stages of alcoholic fermentation. The hydrolysis of the glycosidic terpenes by Sp-ß-gl was studied by gas chromatography, and its ability to efficiently release free terpenols has been demonstrated. The concentrations of geraniol, linalool, α-terpineol, and nerol were significantly increased in treated wines. These results suggest the potential application of this new yeast ß-glucosidase as an aroma-enhancing enzyme in winemaking. PRACTICAL APPLICATION: The search for new ß-glucosidase from yeast sources is important to improve the quality of wines. In this work, an S. pararoseus yeast strain has shown to be capable to produce a ß-glucosidase with suitable combination of properties for functionality in wines and with potential to increase the concentration of free aroma compounds, showing good prospects for an industrial application.