Discovery and Validation of a Urinary Exosome mRNA Signature for the Diagnosis of Human Kidney Transplant Rejection

BackgroundDeveloping a noninvasive clinical test to accurately diagnose kidney allograft rejection is critical to improve allograft outcomes. Urinary exosomes, tiny vesicles released into the urine that carry parent cells’ proteins and nucleic acids, reflect the biologic function of the parent cells within the kidney, including immune cells. Their stability in urine makes them a potentially powerful tool for liquid biopsy and a noninvasive diagnostic biomarker for kidney-transplant rejection.MethodsUsing 192 of 220 urine samples with matched biopsy samples from 175 patients who underwent a clinically indicated kidney-transplant biopsy, we isolated urinary exosomal mRNAs and developed rejection signatures on the basis of differential gene expression. We used crossvalidation to assess the performance of the signatures on multiple data subsets.ResultsAn exosomal mRNA signature discriminated between biopsy samples from patients with all-cause rejection and those with no rejection, yielding an area under the curve (AUC) of 0.93 (95% CI, 0.87 to 0.98), which is significantly better than the current standard of care (increase in eGFR AUC of 0.57; 95% CI, 0.49 to 0.65). The exosome-based signature’s negative predictive value was 93.3% and its positive predictive value was 86.2%. Using the same approach, we identified an additional gene signature that discriminated patients with T cell–mediated rejection from those with antibody-mediated rejection (with an AUC of 0.87; 95% CI, 0.76 to 0.97). This signature’s negative predictive value was 90.6% and its positive predictive value was 77.8%.ConclusionsOur findings show that mRNA signatures derived from urinary exosomes represent a powerful and noninvasive tool to screen for kidney allograft rejection. This finding has the potential to assist clinicians in therapeutic decision making.     

Orthodontic treatment and referral patterns: A survey of pediatric dentists,general practitioners,and orthodontists

Objective

This study aims to assess the orthodontic diagnostic skills, referral patterns, and the perceptions of orthodontic benefits of pediatric and general dentists in comparison with orthodontists.

Materials and methods

Two online surveys were e-mailed to pediatric dentists, general dentistry practitioners, and orthodontists registered as members of the Saudi Dental Society and the Saudi Orthodontic Society. The surveys included questions about the type of orthodontic treatment provided, referral trends, and timing; presumed benefits associated with successful orthodontic treatment; and diagnosis and treatment plans of seven cases representing different malocclusions.

Results

In total, 25 orthodontists, 18 pediatric dentists, and 14 general practitioners completed the survey. Only 38.8% of pediatric dentists and 7.1% of general practitioners reported that they practiced orthodontics clinically. The perceptions of the three groups toward the benefits of orthodontic treatment were comparable in the psychosocial areas. However, the orthodontists perceived significantly lesser effects of orthodontic treatment on the amelioration of temporomandibular disorder (TMD) symptoms. Pediatric dentists tended to rate the need and urgency of treatment higher, while general practitioners tended to rate the need of treatment lower. The selected treatment plans for three early malocclusion cases showed the greatest discrepancies between the orthodontists and the other two groups.

Conclusions

The orthodontists consistently and significantly downplayed the perceived benefit of orthodontic treatment to reduce TMD symptoms. Also, while there was a similarity in the diagnosis, there were notable differences in the proposed treatment approaches, perceived treatment need, and timing of intervention between the three groups of practitioners.     

Prostatic diseases under focus in a university hospital in Eastern Saudi Arabia: A 15-year experience

Objectives:

To explore the spectrum of pathologies diagnosed in prostatic biopsies of Saudi men, and test whether the frequency of diagnosing the malignant fraction has been changed over the last 15 years, and assess the association between chronic inflammation (CI) with both benign prostatic hyperplasia (BPH) and cancer (PCa), and investigate the histological findings of cases presented with acute urinary retention (AUR) clinically.

Methods:

This is a retrospective cohort study including all prostatic biopsies accessed in the files in the Surgical Pathology Laboratory of King Fahd University Hospital, Alkhobar, Kingdom of Saudi Arabia over 15 years (1999-2013) for Saudi men. Age, procedure indication, and final diagnoses were retrieved and slides were reviewed.

Results:

There were 360 cases included in this study with a median age of 65 year-old. The BPH comprised the most (64.7%), while PCa accounted for 89 cases, 13.5% of which were incidental. Most cases of both BPH and PCa were diagnosed in the seventh decade. The frequency of diagnosing PCa did not show a solid rise or fall over time. Chronic inflammation is more related to BPH than to PCa. Only CI showed a significant statistical association with AUR.

Conclusion:

Prostatic diseases show a stable trend over time. While CI is a common dominator for both BPH and PCa, it is associated more with BPH. Among all histological findings, only CI is related to the clinical presentation of AUR.The spectrum of prostatic diseases is a common health concern all over the world. While benign prostatic hyperplasia (BPH) is the most frequent urologic diagnosis in elderly males worldwide, affecting approximately one third of males in their 60’s,1,2 prostatic carcinoma (PCa) is the second most common cause of cancer-related death in males after lung cancer.3 The disease represents clear racial and national differences, with its highest incidence in blacks in the West, and lowest in Asian males.4 Recently, there has been a rising trend for PCa in populations that are thought to be of low risk.5 Furthermore, the relationship between chronic inflammation (CI) and presence and progression of both BPH and PCa is controversial.6-9 On the other hand, acute urinary retention (AUR), which is the most frequent emergency in urology, is mostly developed in a background of BPH.10 Whether other pathologies, such as CI and infarction increase the risk of this complication is debated.11-13 In this work, we retrospectively reviewed the whole spectrum of prostatic pathologies diagnosed in our center during the last 15 years, including prostatic intraepithelial neoplasia (PIN) and atypical small acinar proliferation (ASAP), and study various diseases in relation to age of diagnosis, explore the change in trend of diagnosing PCa over a period witness an increase in screening and gradual westernization of lifestyle, test the relationship between CI and both BPH and PCa, and also, we investigate the role of different biopsy findings in relation to the clinical presentation with AUR.     

Prevalence of cerebral venous thrombosis with the use of oral contraceptive pills during the Holy month of Ramadan

Objectives:To identify the epidemiologic profile of cerebral vein thrombosis (CVT) among fasting women using oral contraceptive pills (OCPs) during the holy month of Ramadan.Methods:This retrospective study was conducted on all patients diagnosed with CVT and using OCPs from records at a tertiary care hospital in Riyadh, Saudi Arabia during 2016-2017. The study participants were categorized into 2 groups (an intermittently fasting group during the holy month of Ramadan and a non-fasting group).Results:Out of 108 female patients with CVT, 36.1% were secondary to OCP, of whom 41% participants were fasting. The most affected site was the transverse sinus. Holocephalic headache was more common amongst fasting group (68.8%) compared to non-fasting group (30.4%) (p=0.025). Dehydration (p=0.003) amongst the fasting group and protein S deficiency (p=0.027) in the non-fasting group were identified as the 2 prominent risk factors. Unfractionated heparin was the most common anticoagulant therapies used during the initiation phase for non-fasting (36.4%) and fasting groups (50%).Conclusion:All women who are using OCP should undergo formal written risk assessments for factors of CVT. Our study suggests that the negative effects of OCPs use might outweigh its benefits; thus, it should be prescribed with caution, more so in fasting patients.     

Vitamin D deficiency in children with bronchial asthma in southern Jordan: a cross-sectional study

ObjectivesTo assess serum 25-hydroxycholecalciferol (25-OH vitamin D) levels in Jordanian children with bronchial asthma, and to examine correlations between 25-OH vitamin D levels and asthma severity and control.MethodsA cross-sectional study was conducted at the Paediatric Chest Clinic, Al-Karak Governmental Hospital, Southern Jordan, between May 2015 and February 2016. Serum 25-hydroxyvitamin D level was determined in children aged 1–14 years diagnosed with bronchial asthma (6–14 years) or recurrent wheezing episodes (<6 years). Asthma severity was determined based on the Global Initiative for Asthma assessment, the Asthma Control Test, and the Childhood Asthma Control Test. Demographic and clinical characteristics were compared between patients with low and normal 25-OH vitamin D levels, and correlations between asthma severity and 25-OH vitamin D level were assessed.ResultsOut of 98 included children, 25-OH vitamin D levels were deficient and insufficient in 41 (41.8%) and 34 (34.7%) children, respectively. Only 23 (23.5%) had sufficient 25-OH vitamin D levels. A significant correlation was found between severity of asthma symptoms and 25-OH vitamin D deficiency.Conclusion25-OH vitamin D deficiency is highly prevalent in Jordanian children with bronchial asthma and correlates significantly with asthma severity.     

PCR-enzyme immunoassay of rDNA in the diagnosis of candidemia and comparison with amplicon detection by agarose gel electrophoresis

We have developed a semi-nested PCR-enzyme immunoassay (snPCR-EIA) for the detection of Candida species in serum specimens, and the sensitivity of amplicon detection was compared with the detection of amplified product by agarose gel electrophoresis (AGE). The universal outer primers amplified the 3' end of 5.8S and the 5' end of 28S rDNA including the internally transcribed spacer 2 (ITS2) in PCR with genomic DNA as template from all the tested Candida species. The biotin-labeled species-specific primers derived from ITS2 from the four commonly encountered Candida species, viz. C. albicans, C. tropicalis, C. parapsilosis and C. glabrata, together with digoxigenin-labeled reverse primer amplified species-specific DNA in the reamplification step of the snPCR. The snPCR-EIA was positive for genomic DNA recovered from 0.06 Candida cells in culture and one organism/ml in spiked serum specimens. Evaluation of snPCR-EIA and snPCR-AGE for specific identification of Candida species with 26 clinical Candida isolates showed 100% concordant results with Vitek and ID32C yeast identification systems. Further evaluation of snPCR-EIA and snPCR-AGE for detection of Candida species in serum samples from culture proven (n = 6) and suspected (n = 10) patients showed concordance with the corresponding species isolated in culture. The serum samples from none of the healthy volunteers (n = 10) were positive for the presence of Candida DNA by snPCR-EIA or snPCR-AGE. Our results show that the snPCR-EIA has the same sensitivity as snPCR-AGE, however, it offers additional advantages of simultaneous testing of a large number of serum samples and avoids the use of ethidium bromide, a potent mutagen. The snPCR-EIA could, therefore, be a method of choice for the diagnosis of candidemia.     

Update on bereavement research: evidence-based guidelines for the diagnosis and treatment of complicated bereavement

The past decade has witnessed considerable growth in the evidence-base from which clinical recommendations for bereavement care can be made. Research now provides guidance to assist clinicians in: a) recognizing differences between complicated and uncomplicated bereavement reactions, b) identifying risk factors that may make certain individuals more vulnerable to bereavement-related complications, c) appreciating and monitoring for potential adverse outcomes associated with bereavement and d) taking actions to prevent or minimize maladjustment to the loss. In this article we distinguish between the course of normal grief and abnormally prolonged, or complicated grief; clarify distinctions between Complicated Grief Disorder and other mental disorders secondary to bereavement; review outcomes associated with Complicated Grief Disorder; describe research on resilience in bereavement; present findings on stigmatization and the use of mental health services among recently bereaved persons; and summarize where the field is with respect to establishing the efficacy and effectiveness of bereavement interventions. Promising new psychotherapies for Complicated Grief Disorder have shown clinical efficacy. Nevertheless, further research is needed to enhance the detection of vulnerable bereaved persons, to promote resilience following significant interpersonal loss, and to tailor interventions to address the attachment issues that lie at the heart of this disorder.