Unraveling the expression of microRNA-27a* & NKG2D in peripheral blood mononuclear cells and natural killer cells of pediatric systemic lupus erythematosus patients.

BACKGROUND AND AIM: The activity of natural killer (NK) cells is known to be decreased in systemic lupus erythematosus (SLE) patients. Nevertheless, the exact contribution of NK cells in the pathogenesis of SLE is still inconclusive. MicroRNAs (miRNAs), are small noncoding RNA molecules that play a fundamental role in regulating NK cell function. The objective of this study was to investigate the expression of miRNAs that might potentially target an essential activating receptor, NKG2D in peripheral blood mononuclear cells (PBMCs) and NK cells of SLE patients. METHODS: In silico analysis revealed miR-27a* to potentially target NKG2D messenger RNA (mRNA), hence PBMCs and NK cells were isolated from blood samples of SLE patients and healthy controls. Next, the cells were transfected using mimics and antagomirs, after which miRNA/mRNA were quantified using real time quantitative reverse transcription polymerase chain reaction. RESULTS: The results of this study showed that miR-27a* is overexpressed in the PBMCs and NK cells of SLE patients. In contrast, NKG2D was found to be downregulated in PBMCs and NK cells of SLE patients. Forcing the expression of miR-27a* in PBMCs and NK cells enhances the expression of NKG2D in SLE patients. Furthermore, the ligand of NKG2D, ULBP2, was found to be downregulated in the PBMCs of SLE patients. CONCLUSION: The altered expression of the triad, miR-27a* as well as NKG2D and ULBP2, is thought to be characteristic for NK cells in SLE patients. Hence, the ability of miR-27a* to alter the expression of NKG2D may provide a new groundwork for understanding the role of miRNAs in NK cells of SLE patients.

A mini review of qRT-rtPCR technology application in uncovering the mechanism of food allergy and in the search for novel interventions.

This mini review targets the inclusion of recent selected citations, between the year 2006 and 2012, that implement the qRT-rtPCR technology in their experimental designs, targeting the uncovering of the mechanism of food allergy. In addition, this same technology was implemented in specific experimental designs, aiming at finding novel nutritional, herbal medicine, and tolerance interventions against food allergy. The approach of using qRT-rtPCR technology helped in studying the dynamics of transcription of cytokines and chemokines in intestinal dendritic cells of the experimental animals during the allergic reaction to food. The suppression of transcription of specific cytokines or chemokines by nutritional, herbal medicine, and tolerance interventions was instrumental in the search for finding novel remedies for this health condition, that was traditionally managed by avoidance of offending foods in the diet.

Optimization of Saanen sperm genes amplification: evaluation of standardized protocols in genetically uncharacterized rural goats reared under a subtropical environment.

The purpose of this research is to optimize quantitatively the amplification of specific sperm genes in reference genomically characterized Saanen goat and to evaluate the standardized protocols applicability on sperms of uncharacterized genome of rural goats reared under subtropical environment for inclusion in future selection programs. The optimization of the protocols in Saanen sperms included three production genes (growth hormone (GH) exons 2, 3, and 4, αS1-casein (CSN1S1), and α-lactalbumin) and two health genes (MHC class II DRB and prion (PrP)). The optimization was based on varying the primers concentrations and the inclusion of a PCR cosolvent (Triton X). The impact of the studied variables on statistically significant increase in the yield of amplicons was noticed in four out of five (80%) optimized protocols, namely in those related to GH, CSN1S1, α-lactalbumin, and PrP genes (P < 0.05). There was no significant difference in the yield of amplicons related to MHC class II DRB gene, regardless of the variables used (P > 0.05). The applicability of the optimized protocols of Saanen sperm genes on amplification of uncharacterized rural goat sperms revealed a 100% success in tested individuals for amplification of GH, CSN1S1, α-lactalbumin, and MHC class II DRB genes and a 75% success for the PrP gene. The significant success in applicability of the Saanen quantitatively optimized protocols to other uncharacterized genome of rural goats allows for their inclusion in future selection, targeting the sustainability of this farming system in a subtropical environment and the improvement of the farmers livelihood.

MR diagnosis of rotator cuff tears of the shoulder: value of using T2-weighted fat-saturated images.

OBJECTIVE: The purpose of this study was to compare the use of fat-saturated T2-weighted spin-echo MR imaging with that of conventional T2-weighted spin-echo MR imaging to detect full- and partial-thickness tears of the rotator cuff of the shoulder, using arthroscopy as the standard for the diagnosis. SUBJECTS AND METHODS: Forty-nine consecutive patients with shoulder pain who had both MR imaging and arthroscopy of their affected shoulders were studied. The study group consisted of 10 patients with full-thickness tears of the rotator cuff, 20 with partial-thickness tears of the rotator cuff, and 19 with intact rotator cuff tendons proved arthroscopically. All patients had T1-weighted, conventional T2-weighted, and fat-saturated T2-weighted MR images obtained in a plane slightly oblique to the coronal plane along the long axis of the supraspinous tendon. The images were divided into two sets, one including both T1-weighted and conventional T2-weighted images, the other including T1-weighted and fat-saturated T2-weighted images. Two musculoskeletal radiologists reviewed each set of MR images in a blinded fashion. The reviewers were asked to decide whether the rotator cuff showed a full-thickness tear, a partial-thickness tear, or no tear on each set of images. In the case of partial-thickness tears, the reviewers were asked to indicate which surface (bursal or joint) of the rotator cuff was affected. These data were correlated with the arthroscopic findings and with each other using McNemar and kappa analysis. RESULTS: Detection of full-thickness tears was excellent for both reviewers using either imaging technique. However, when the fat-saturated technique was used, the sensitivity increased significantly, from 80% to 100%. Detection of partial-thickness tears was poor with conventional spin-echo MR imaging (15%). Although significantly improved when fat saturation was used (35%), detection rates were still lower than rates reported in the literature. Identification of the torn surface of the rotator cuff was correct in 50% of cases in which partial-thickness tears were successfully identified. Fat saturation decreased the specificity of identification of both partial and full-thickness tears of the rotator cuff but significantly increased the specificity with which intact rotator cuffs were identified. CONCLUSION: Use of the fat-saturation technique improved detection of both full-thickness and partial-thickness tears of the rotator cuff on MR images compared with standard spin-echo imaging techniques. Despite this improvement, detection of partial-thickness tears was poor with both techniques. Furthermore, correct identification of which surface of the rotator cuff was torn in patients with partial tears was nearly random.

MR imaging of the adrenal glands.

MR imaging provides an excellent morphologic depiction of normal and abnormal adrenal glands. Although CT scanning remains the primary imaging technique for detecting adrenal lesions, MR imaging is being used increasingly for characterization of adrenal masses. Chemical shift imaging is the most recent MR technique to be evaluated for distinguishing benign from malignant masses, and it appears to be the most reliable; however, further validation of this technique is necessary. In most clinical situations, adrenal lesions thought to be malignant on the basis of MR criteria should still be biopsied. MR imaging is also a useful problem-solving technique after equivocal CT scanning or sonography.

Ultrasonographic evaluation of the biliary tree in patients with atypical chest pain.

We studied the use of gallbladder ultrasonography in the evaluation of atypical chest pain (ACP). During an 8-month period, 52 patients were referred for biliary ultrasonography with a chief complaint of chest pain. Twelve (32.1%) had cholelithiasis discovered by ultrasonography. Four of these 12 (33.3%) underwent subsequent cholecystectomy with complete relief of symptoms. None of the patients with normal ultrasonography for whom follow-up was available (32) underwent subsequent cholecystectomy. We found no statistical difference in the prevalence of cholelithiasis between the ACP group and 85 asymptomatic controls. Clinical evidence confirms the concept that ultrasonography may play an important role in the evaluation of ACP.

Preoperative evaluation of intraabdominal tumors in children: gradient-recalled echo vs spin-echo MR imaging.

OBJECTIVE: The purpose of this study was to compare gradient-recalled echo (GRE) with spin-echo (SE) MR imaging in the assessment of vessel patency and tumor margins in children with intraabdominal tumors. MATERIALS AND METHODS: Findings on GRE and SE images in 10 children with abdominal neoplasms were retrospectively evaluated and compared with pathologic findings or results of CT studies. RESULTS: Detection of tumor thrombus was 100% with GRE sequences and 75% with SE sequences. The SE sequences yielded two false-positive diagnoses of tumor thrombosis, whereas the GRE sequences yielded no false-positive or false-negative diagnoses. In patients with extrinsic compression of the inferior vena cava by tumor, GRE images were consistently better than SE images for confirming vessel patency. GRE images also were superior to SE images at showing flow in mesenteric vessels, renal arteries, and renal veins. However, GRE images displayed tumor margins less well than SE images did, and they were also more susceptible to ghosting artifacts. CONCLUSION: Our results suggest that the GRE sequence is a useful adjunct to SE sequences for evaluating vascular involvement in children with abdominal tumors.

Gadopentetate dimeglumine-enhanced MR imaging of the postoperative lumbar spine: comparison of fat-suppressed and conventional T1-weighted images.

OBJECTIVE: Gadopentetate dimeglumine-enhanced T1-weighted MR imaging has been used to distinguish epidural fibrosis from recurrent disk herniation in the postoperative lumbar spine. Potential interpretative difficulties relate to detection of subtle contrast enhancement and distinction of enhanced tissue from surrounding epidural fat. The objective of this study was to evaluate the utility of fat-suppressed T1-weighted imaging in improving the depiction of postoperative abnormalities. SUBJECTS AND METHODS: Forty-one patients with recurrent signs and symptoms after previous laminectomy and diskectomy were examined with conventional and fat-suppressed T1-weighted imaging before and after IV administration of gadopentetate dimeglumine. MR images were analyzed for the presence or absence of an epidural soft-tissue mass located anterior and/or lateral to the thecal sac. When such masses were identified, their contrast enhancement behavior was characterized according to the volume, intensity, and pattern of enhancement. Epidural fibrosis was diagnosed by using established MR imaging criteria. In patients with evidence of recurrent disk herniation, diagnosis was based on surgery in six patients and on MR imaging findings in the remainder. RESULTS: Soft-tissue masses were present at 32 of the 43 levels operated on, representing epidural fibrosis (n = 21) or disk herniation (n = 11). Fat suppression improved depiction of contrast enhancement associated with sites of epidural fibrosis. The extent, intensity, and homogeneity of enhancement was greater with fat suppression than with conventional imaging, although the ability to distinguish epidural fibrosis from recurrent disk herniation was not improved. CONCLUSION: Use of the fat-suppression technique results in increased relative intensity and homogeneity of contrast enhancement associated with sites of epidural fibrosis. However, it does not result in significant improvement in the ability to differentiate fibrotic tissue from recurrent disk herniation. Fat suppression does not appear to be indicated for routine use in imaging of the postoperative lumbar spine, but may be useful as a problem-solving tool in selected patients.

CT of focal pulmonary masses in childhood.

Focal pulmonary masses in children encompass a spectrum of conditions including congenital lesions, inflammatory masses, hematomas, and benign and malignant tumors. Congenital masses include bronchial atresia, sequestration, cystic adenomatoid malformation, and pulmonary vascular anomalies. Inflammatory masses most commonly result from infection and include pulmonary abscess, granulomas, and postinflammatory pseudotumors. Blunt trauma can cause a hematoma, which decreases in size on serial radiographs. Pulmonary neoplasms may be benign such as papilloma, hamartoma, and bronchial carcinoid tumor (low-grade malignancy), or they may be malignant as in sarcoma, carcinoma, and pulmonary blastoma. Because computed tomography (CT) is the most sensitive technique in detecting and helping characterize parenchymal disease, it has become the procedure of choice for further investigation of lesions seen or suggested on plain chest radiographs. Understanding the CT appearance of these lesions can allow an accurate diagnosis and optimize management of the patient's condition.

Ultrasound evaluation of the common duct in symptomatic and asymptomatic patients.

We studied common duct (CD) diameter using real time ultrasound in 551 patients. The patients were divided into two groups: those referred for evaluation of the biliary tree [biliary-referred group (BRG)], and those referred for a problem unrelated to potential biliary tree pathology [non-biliary-referred group (NBRG)]. Potentially obstructed CDs were excluded from analysis. The CD diameter in the NBRG patients was found to increase after cholecystectomy (p less than 0.0003), but not in patients in this group with intact gallbladders and cholelithiasis. Dilation was observed in the BRG patients with either cholelithiasis (p less than 6 x 10(-7)) or cholecystectomy (p less than 4 x 10(-10)) compared to patients with intact gallbladders without cholelithiasis. To our knowledge, the former phenomenon has not been described in the literature. Our NBRG data suggest that the normal CD diameter should be set at 5 mm and 8 mm, respectively, for patients with intact gallbladders and patients post-cholecystectomy. No correlation was found between duct size and sex, age, or weight.

Postgastrostomy tube deformity on upper GI series.

Three cases are presented in which a focal concave deformity occurred along the greater curvature of the stomach on upper gastrointestinal (GI) series. These patients all had recent removal of a surgically placed gastrostomy tube from a similar location. This deformity appears to be related, at least in part, to invaginated gastric mucosa intentionally produced during surgical gastrostomy tube placement. This association and appearance should be noted as it may mimic other lesions.