TMEM106B is a receptor mediating ACE2-independent SARS-CoV-2 cell entry.

SARS-CoV-2 is associated with broad tissue tropism, a characteristic often determined by the availability of entry receptors on host cells. Here, we show that TMEM106B, a lysosomal transmembrane protein, can serve as an alternative receptor for SARS-CoV-2 entry into angiotensin-converting enzyme 2 (ACE2)-negative cells. Spike substitution E484D increased TMEM106B binding, thereby enhancing TMEM106B-mediated entry. TMEM106B-specific monoclonal antibodies blocked SARS-CoV-2 infection, demonstrating a role of TMEM106B in viral entry. Using X-ray crystallography, cryogenic electron microscopy (cryo-EM), and hydrogen-deuterium exchange mass spectrometry (HDX-MS), we show that the luminal domain (LD) of TMEM106B engages the receptor-binding motif of SARS-CoV-2 spike. Finally, we show that TMEM106B promotes spike-mediated syncytium formation, suggesting a role of TMEM106B in viral fusion. Together, our findings identify an ACE2-independent SARS-CoV-2 infection mechanism that involves cooperative interactions with the receptors heparan sulfate and TMEM106B.

TGF-ß induces the expression of the adaptor Ndfip1 to silence IL-4 production during iTreg cell differentiation.

Mice deficient in the adaptor Ndfip1 develop inflammation at sites of environmental antigen exposure. We show here that such mice had fewer inducible regulatory T cells (iT(reg) cells). In vitro, Ndfip1-deficient T cells expressed normal amounts of the transcription factor Foxp3 during the first 48 h of iT(reg) cell differentiation; however, this expression was not sustained. Abortive Foxp3 expression was caused by production of interleukin 4 (IL-4) by Ndfip1(-/-) cells. We found that Ndfip1 expression was transiently upregulated during iT(reg) cell differentiation in a manner dependent on transforming growth factor-ß (TGF-ß). Once expressed, Ndfip1 promoted degradation of the transcription factor JunB mediated by the E3 ubiquitin ligase Itch, thus preventing IL-4 production. On the basis of our data, we propose that TGF-ß signaling induces Ndfip1 expression to silence IL-4 production, thus permitting iT(reg) cell differentiation.

Correlation Between Hemoglobin Glycation Index Measured by Continuous Glucose Monitoring With Complications in Type 1 Diabetes.

OBJECTIVE: HbA1C is the "gold standard" parameter to evaluate glycemic control in diabetes; however, its correlation with mean glucose is not always perfect. The objective of this study was to correlate continuous glucose monitoring (CGM)-derived hemoglobin glycation index (HGI) with microvascular complications. METHODS: We conducted a cross-sectional study including permanent users of CGM with type 1 diabetes mellitus or latent autoimmune diabetes of the adult. HGI was estimated, and presence of microvascular complications was compared in subgroups with high or low HGI. A logistic regression analysis to assess the contribution of high HGI to chronic kidney disease (CKD) was performed. RESULTS: In total, 52 participants who were aged 39.7 ± 14.7 years, with 73.1% women and 15.5 years (IQR, 7.5-29 years) since diagnosis, were included; 32.7% recorded diabetic retinopathy, 25% CKD, and 19.2% neuropathy. The median HbA1C was 7.6% (60 mmol/mol) and glucose management indicator (GMI) 7.0% (53 mmol/mol). The average HGI was 0.55% ± 0.66%. The measured HbA1C was higher in the group with high HGI (8.1% [65 mmol/mol] vs 6.9% [52 mmol/mol]; P < .001), whereas GMI (7.0% [53 mmol/mol] vs 7.0% [53 mmol/mol]; P = .495) and mean glucose were similar in both groups (153 mg/dL vs 153 mg/dL; P = .564). In the high HGI group, higher occurrence of CKD (P = .016) and neuropathy were observed (P = .025). High HGI was associated with increased risk of CKD (odds ratio [OR]: 5.05; 95% CI: 1.02-24.8; P = .04) after adjusting for time since diagnosis (OR: 1.09; 95% CI: 1.02-1.16; P = .008). CONCLUSION: High HGI measured by CGM may be a useful marker for increased risk of microvascular diabetic complications.

Role of Bladder Functional Testing Prior to Surgeries for Benign Prostatic Obstruction.

PURPOSE OF REVIEW: There is no consensus on preoperative functional testing prior to surgeries for benign prostatic obstruction causing lower urinary tract symptoms (LUTS). RECENT FINDINGS: Surgical management offers definite benefits, but the results are not always satisfactory. The urodynamic study (UDS) is the gold standard for assessing bladder outlet obstruction (BOO) which is the best predictor of surgical success. Yet, it is not recommended by our urologic societies as standard testing prior to surgery. In this narrative review of the literature, we report recent findings and controversies regarding the benefits and downside of UDS, and the use of other less-invasive approaches to achieve this goal. The lack of strong evidence for or against performing UDS was surprising. Prospective UDS data may not predict surgical outcomes if there is no consensus on criteria that directs surgical intervention. However, confirming the presence of BOO and characterizing the bladder function to identify detrusor over- and underactivity may help counselling and setting patient's post-operative expectations. Urocuff, a non-invasive testing offers promising results to address this problem with a less-invasive assessment of BOO. We emphasize better pre-operative characterization of patients to confirm BOO and better define subgroups to improve surgical decision-making.

Role of the arts in the life and mental health of young people that participate in artistic organizations in Colombia: a qualitative study.

BACKGROUND: Adolescents and young adults are vulnerable to developing mental distress. However, evidence suggests that more than half of the young people with symptoms of depression and anxiety overcome their distress within a year. However, there is little research on the exact resources that young people use and help them to recover. The aim of this study was to explore how arts activities can support the recovery of young people engaged with arts organizations in Bogota. METHODS: We recruited 38 participants from two arts organizations in Bogotá and conducted six focus groups embedded within artistic workshops. The type of activities in the workshops varied reflecting the different teaching methods of the two organizations. The focus group discussions were recorded and analyzed using thematic analysis. RESULTS: Five themes explained how arts activities can help young people participating in artistic organizations to overcome mental distress: i) allowing the expression of emotions; ii) helping to manage and transform emotions; iii) distracting from problems; iv) facilitating social support and relationships; and v) contributing to the identity of young people. CONCLUSIONS: For young people who participate in artistic organizations, the arts are a resource for overcoming negative emotions such as anxiety, depression, and sadness. The beneficial role of arts activities includes different process of managing, expressing, and distracting from distress, and it differs depending on whether arts are perceived as a professional vocation or a hobby.

The genomic basis of army ant chemosensory adaptations.

The evolution of mass raiding has allowed army ants to become dominant arthropod predators in the tropics. Although a century of research has led to many discoveries about behavioural, morphological and physiological adaptations in army ants, almost nothing is known about the molecular basis of army ant biology. Here we report the genome of the iconic New World army ant Eciton burchellii, and show that it is unusually compact, with a reduced gene complement relative to other ants. In contrast to this overall reduction, a particular gene subfamily (9-exon ORs) expressed predominantly in female antennae is expanded. This subfamily has previously been linked to the recognition of hydrocarbons, key olfactory cues used in insect communication and prey discrimination. Confocal microscopy of the brain showed a corresponding expansion in a putative hydrocarbon response centre within the antennal lobe, while scanning electron microscopy of the antenna revealed a particularly high density of hydrocarbon-sensitive sensory hairs. E. burchellii shares these features with its predatory and more cryptic relative, the clonal raider ant. By integrating genomic, transcriptomic and anatomical analyses in a comparative context, our work thus provides evidence that army ants and their relatives possess a suite of modifications in the chemosensory system that may be involved in behavioural coordination and prey selection during social predation. It also lays the groundwork for future studies of army ant biology at the molecular level.

Effectiveness of convective water vapor energy therapy versus prostatic urethral lift for symptomatic benign prostatic hyperplasia: a systematic review and indirect comparison.

PURPOSE: To synthesize the evidence from randomized controlled trials of prostatic urethral lift (PUL) and convective water vapor thermal energy therapy (WAVE) for minimally invasive treatment of men with benign prostatic hyperplasia. METHODS: A systematic search of databases was performed to identify trials comparing WAVE or PUL to either an active or sham surgery control in subjects with symptomatic benign prostatic obstruction. A controlled indirect treatment comparison based on the approach of Bucher was performed for outcomes including International Prostate Symptom Score and maximum urinary flow rate (Qmax). The durability of treatment response was assessed by life-table analysis of freedom from retreatment through 4 years. RESULTS: Two multicenter sham-controlled trials (Rezum II Study, NCT01912339: LIFT Study, NCT01294150) were identified. The trials employed a common sham procedure and were similar with respect to their designs and subjects' baseline characteristics. Comparisons on the treatment effect in excess of sham response found non-significant differences between WAVE and PUL for symptom score [mean difference (MD): - 1.7 points; 95% confidence interval (CI): - 4.8, 1.4] but Qmax improvements favored WAVE [MD: 3.4 ml/sec; CI: 1.2, 5.6]. The proportion free of retreatment through 4 years was 89.1% for WAVE versus 75.4% for PUL [log-rank P = 0.004]. CONCLUSIONS: PUL and WAVE provide similar subjective improvements but flow-rate improvement and durability of response seem greater for WAVE. The confirmation of these findings in a randomized trial is warranted.

HLA-B Allele, Genotype, and Haplotype Frequencies in a Group of Healthy Individuals in Colombia.

BACKGROUND: The sequencing of alleles of the HLA-B, a human leukocyte antigen (HLA) class I gene, was established as the most polymorphic of chromosome 6 and of the entire human genome. In this locus, the HLA-B*27 allele is highly polymorphic and has clinical relevance. Literature about the subtypes and singular frequency of these alleles in Colombia's healthy population is scarce. OBJECTIVE: The aim of this study was to establish the HLA-B allele, genotype, and haplotype frequencies in a healthy Colombian population and analyze their association with the sex and geographical distribution of the individuals studied. METHODS: This is a nonexperimental and descriptive study. The data from whole-blood samples whose HLA genes were genotyped by protocol with the Luminex 100/200 xMAP technology were evaluated. HLA-B*27 positivity was confirmed by the new-generation sequencing technology. The associations between the HLA-B alleles and demographic variables were evaluated by χ2 and Fisher exact tests. RESULTS: Twenty-seven HLA-B genotypes were identified in 255 individuals, with the highest frequencies for HLA-B*35 (44.7%), B*40 (19.6%), and B*44 (16.8%). Additionally, 89 HLA-B alleles were found; the most common were HLA-B*35:01 (6.7%) and B*40:02 (6.5%). Nine individuals tested positive for the HLA-B*27 allele with genotype and allele frequencies of 3.5% and 1.8%, respectively; the HLA-B*27:05:02 subtype predominated. CONCLUSIONS: Here, we report the most common HLA-B allele, genotype, and haplotype frequencies in a healthy Colombian population group and analyzed their association with the sex and geographical distribution of the individuals studied. Results for the HLA-B*27 allele confirm racial mixing in Colombia with a high degree of Caucasian influence, as well as the repopulation of Colombia's central region, attributed to the migration phenomena. Results agree with data published in Colombia that was obtained from cord blood samples.

Type 1 Diabetes Care in Mexico: An Analysis of the RENACED-DT1 National Registry.

BACKGROUND: Information regarding diagnosis, treatment, and follow-up of patients with type 1 diabetes (PWT1D) in Mexico is limited. We developed an on-line platform Registro Nacional de Pacientes con Diabetes Tipo 1 (RENACED-DT1). OBJECTIVE: The objective of the study was to describe the characteristics and healthcare of PWT1D registered in RENACED-DT1. METHODS: Analyses of 965 PWT1D from July 2014 to January 2018 in different endocrinology clinics around Mexico. RESULTS: Sixty-one percent were female with median age of 21 years, age at diagnosis 11 years, and disease duration at inclusion 8.2 years. Treatment regimen was basal-bolus in 61% and insulin-pumps in 21% (mainly in the private sector); 33.3% with self-monitoring of blood-glucose (SMBG) ≥4 times/day. Mean HbA1c at last follow-up was 8.7 ± 2.1% (72±23 mmol/mol), 18% had HbA1c < 7% (53 mmol/mol), and 35% > 9% (75 mmol/mol). SMBG ≥ 4 times/day was associated with HbA1c < 7%. Time since diagnosis > 10 years, female sex, BMI ≥ 30 kg/m2, SMBG < 4 times/day, and any hypoglycemia were associated with microvascular complications (p < 0.05). CONCLUSIONS: Percentage of patients achieving HbA1c < 7% is low; increased blood glucose monitoring is associated with better glycemic control. The achievement of optimal glycemic control must be increased to reduce the incidence of chronic complications and improve quality of life in PWT1D.

Cessation of Ureteral Colic Does Not Necessarily Mean that a Ureteral Stone Has Been Expelled.

PURPOSE: We evaluated whether cessation of renal colic is consistent with an expelled ureteral stone or whether imaging may be indicated even in the absence of symptoms. MATERIALS AND METHODS: We performed a retrospective study of patients who presented to our institution with acute renal colic and ureteral stone, and were subsequently evaluated at a followup visit where they reported complete cessation of pain for at least 72 hours. RESULTS: Study inclusion criteria were met by 52 patients, who reported no pain for at least 72 hours at the time of the followup visit. A persistent ureteral stone was demonstrated in 14 of the 52 patients (26%) although they denied any associated symptoms. Multivariate logistic regression did not show an association between stone size or location and the likelihood of passage in this cohort. CONCLUSIONS: Cessation of pain was associated with ureteral stone passage in almost 75% of this study cohort but 26% of patients still had persistent ureteral stones. We recommend routine followup imaging in all patients with ureteral stones to document stone passage and avoid the risks of silent ureteral obstruction.

Forest resilience to drought varies across biomes.

Forecasted increase drought frequency and severity may drive worldwide declines in forest productivity. Species-level responses to a drier world are likely to be influenced by their functional traits. Here, we analyse forest resilience to drought using an extensive network of tree-ring width data and satellite imagery. We compiled proxies of forest growth and productivity (TRWi, absolutely dated ring-width indices; NDVI, Normalized Difference Vegetation Index) for 11 tree species and 502 forests in Spain corresponding to Mediterranean, temperate, and continental biomes. Four different components of forest resilience to drought were calculated based on TRWi and NDVI data before, during, and after four major droughts (1986, 1994-1995, 1999, and 2005), and pointed out that TRWi data were more sensitive metrics of forest resilience to drought than NDVI data. Resilience was related to both drought severity and forest composition. Evergreen gymnosperms dominating semi-arid Mediterranean forests showed the lowest resistance to drought, but higher recovery than deciduous angiosperms dominating humid temperate forests. Moreover, semi-arid gymnosperm forests presented a negative temporal trend in the resistance to drought, but this pattern was absent in continental and temperate forests. Although gymnosperms in dry Mediterranean forests showed a faster recovery after drought, their recovery potential could be constrained if droughts become more frequent. Conversely, angiosperms and gymnosperms inhabiting temperate and continental sites might have problems to recover after more intense droughts since they resist drought but are less able to recover afterwards.

Eating behaviors and emotional distress are predicted by treatment and adverse outcome in patients with type 2 diabetes.

Depression and anxiety are hightly prevalent among patients with Type 2 Diabetes (T2D), however not commonly related to outcomes, treatment and comorbidities. Eating behaviors could also have an implication. To evaluate the relation between mood and eating behaviors with demographic, physical, treatment, biochemical profiles and chronic comorbidities in T2D we conducted this exploratory cross sectional study in a population from Mexico. Hospital Anxiety and Depression Scale and the Three Factor Eating Questionnaire Revised 21 (TFEQ-R21) were correlated with age, gender, blood pressure, treatment and comorbidities (nephropathy, neuropathy, retinopathy and cardiopathy). Multi-linear regression models and 2k factorial analyses were conducted. Sixty-one patients (31 male) 55 years old (SD 13) with at least 5 years of T2D were included. Anxiety correlated with depression (r = 0.25, p < 0.05). Gender (women) (std B 0.026, p < 0.001), insulin therapy (std B 0.3, p = 0.11), systolic blood pressure (std B 0.263, p = 0.02) and cardiovascular disease (std B 0.232, p = 0.035) predicted depression. Insulin therapy had a positive effect in cognitive restraint. (std B 0.32, p = 0.001). Age (std B 0.37, p = 0.003) and systolic blood pressure (std B 0.237, p = 0.048) predicted positively emotional eating (not previously published). 2k factorial analyses proved additive interaction between complications and insulin therapy with depression. This study supports the relation between mood disorders and eating behaviors with demographic, physical, treatment, biochemical profiles and chronic comorbidities in T2D and the additive interaction between factors and mood disorders.

Is Hydronephrosis on Ultrasound Predictive of Ureterolithiasis in Patients with Renal Colic?

PURPOSE: Renal ultrasound accurately identifies hydronephrosis but it is less sensitive than computerized tomography for the detection of ureterolithiasis. We investigated whether the presence of hydronephrosis on ultrasound was associated with a ureteral stone in patients who underwent both ultrasound and computerized tomography during the evaluation of acute renal colic. MATERIALS AND METHODS: We retrospectively reviewed the records of patients from 3 institutions who were evaluated for acute renal colic by both ultrasound and computerized tomography between 2012 and 2015. Patients were included in analysis if ultrasound and computerized tomography were performed on the same day. The presence of ureterolithiasis, stone location and hydronephrosis was reviewed and compared between imaging modalities. RESULTS: Ureteral stones were present in 85 of 144 patients. Ultrasound identified hydronephrosis in 89.8% of patients and a ureteral stone in 25.9%. Computerized tomography identified hydronephrosis in 91.8% of patients and a ureteral stone in 98.8%. In 75.0% of cases the presence or absence of hydronephrosis on ultrasound correctly predicted the presence or absence of a ureteral stone on computerized tomography. Hydronephrosis on ultrasound had a positive predictive value of 0.77 for the presence of a ureteral stone and a negative predictive value of 0.71 for the absence of a ureteral stone. CONCLUSIONS: Hydronephrosis on ultrasound did not accurately predict the presence or absence of a ureteral stone on computerized tomography in 25.0% of the patients in this study. Ultrasound is an important tool for evaluating hydronephrosis associated with renal colic but patients may benefit from other studies to confirm the presence or absence of ureteral stones.

Predicting ureteral stones in emergency department patients with flank pain: an external validation of the STONE score.

PURPOSE: The ability to objectively predict which emergency department patients are likely to have a ureteral stone may aid in cost-effectiveness and patient-centered diagnostic imaging decisions. We performed an external validation of the STONE score, a clinical prediction rule for the presence of uncomplicated ureteral stones in emergency department patients developed at Yale University School of Medicine. METHODS: Five hundred thirty-six (536) consecutive patients evaluated in an urban tertiary care emergency department for the possible diagnosis of ureteral stone were retrospectively reviewed. The STONE score uses five factors (gender, duration of pain, race, nausea/vomiting, erythrocytes on urine dipstick) to categorize patients into low, medium, and high probability of having a ureteral stone. The total STONE score risk is 0-13 and divided into three groups: low risk = 0-5, moderate risk = 6-9, and high risk = 10-13. RESULTS: Of the 536 patients evaluated for suspected ureteral stone, 257 (47.8 %) had a ureteral stone. Mean patient age was 45.9 years (SD 16.3), and gender distribution was 43.9 % female and 56.1 % male. Distribution of STONE score risk was 24.1 % low, 48.1 % moderate, and 27.7 % high. Diagnosis of ureteral stone by STONE score risk was 14 % for low-risk group, 48.3 % for moderate-risk group, and 75.8 % for high-risk group. This distribution is consistent with internal validation at Yale University School of Medicine, where values were 8.3-9.2 % for low risk, 51.3-51.6 % for moderate risk, and 88.6-89.6 % for high risk. CONCLUSIONS: Our study validates the use of the STONE clinical score to categorize patients as low, moderate, and high risk for ureteral stone. This could help guide development of clinical decision rules for diagnostic studies and imaging in the future.

Can ureteral stones cause pain without causing hydronephrosis?

INTRODUCTION: While computerized tomography (CT) is the gold standard for diagnosis of ureterolithiasis, ultrasound is a less costly and radiation-free alternative which is commonly used to evaluate patients with ureteral colic. The purpose of this study was to evaluate the frequency with which patients with ureteral stones and renal colic demonstrate hydronephrosis in order to better understand the evaluation of these patients. METHODS: Two hundred and forty-eight consecutive patients presenting with ureteral colic and diagnosed with a single unilateral ureteral stone on CT scan in an urban tertiary care emergency department were retrospectively reviewed. Radiology reports were reviewed for stone size, diagnosis, and degree of hydronephrosis. RESULTS: Of the 248 patients evaluated for suspected ureteral stone, 221 (89.1 %) demonstrated any hydronephrosis, while 27 (10.9 %) did not. Hydronephrosis grade, available in 194 patients, was as follows: mild-70.6 %, moderate-27.8 %, and severe-1.5 %. Mean patient age was 47.0 years (SD 15.5), gender distribution was 35.9 % female and 64.1 % male, and mean stone axial diameter was 4.1 mm (SD 2.4). Stone location was as follows: ureteropelvic junction-4.1 %, proximal ureter-21 %, distal ureter-24.9 %, and ureterovesical junction-47.1 %. Axial stone diameter and coronal length (craniocaudal) were both significant predictors of degree of hydronephrosis (ANOVA, p < 0.001 for both). Age (ANOVA, p = NS), stone location (Chi square, p = NS), and gender (Chi square, p = NS) were not associated with degree of hydronephrosis. CONCLUSIONS: In patients with ureteral stones and colic, nearly 11 % do not demonstrate any hydronephrosis and a majority (nearly 71 %) will demonstrate only mild hydronephrosis. Stone diameter appears to be related to degree of hydronephrosis, whereas age, gender, and stone location are not. The lower incidence of hydronephrosis for small stones causing renal colic may explain the lower diagnostic accuracy of ultrasound when compared to CT for detecting ureteral stones.

Requirement for MyD88 signaling in B cells and dendritic cells for germinal center anti-nuclear antibody production in Lyn-deficient mice.

The intracellular tyrosine kinase Lyn mediates inhibitory receptor function in B cells and myeloid cells, and Lyn(-/-) mice spontaneously develop an autoimmune and inflammatory disease that closely resembles human systemic lupus erythematosus. TLR-signaling pathways have been implicated in the production of anti-nuclear Abs in systemic lupus erythematosus and mouse models of it. We used a conditional allele of Myd88 to determine whether the autoimmunity of Lyn(-/-) mice is dependent on TLR/MyD88 signaling in B cells and/or in dendritic cells (DCs). The production of IgG anti-nuclear Abs, as well as the deposition of these Abs in the glomeruli of the kidneys, leading to glomerulonephritis in Lyn(-/-) mice, were completely abolished by selective deletion of Myd88 in B cells, and autoantibody production and glomerulonephritis were delayed or decreased by deletion of Myd88 in DCs. The reduced autoantibody production in mice lacking MyD88 in B cells or DCs was accompanied by a dramatic decrease in the spontaneous germinal center (GC) response, suggesting that autoantibodies in Lyn(-/-) mice may depend on GC responses. Consistent with this view, IgG anti-nuclear Abs were absent if T cells were deleted (TCRß(-/-) TCRδ(-/-) mice) or if T cells were unable to contribute to GC responses as the result of mutation of the adaptor molecule SAP. Thus, the autoimmunity of Lyn(-/-) mice was dependent on T cells and on TLR/MyD88 signaling in B cells and in DCs, supporting a model in which DC hyperactivity combines with defects in tolerance in B cells to lead to a T cell-dependent systemic autoimmunity in Lyn(-/-) mice.

Diagnostic and management considerations for nephrolithiasis in the gravid patient.

Renal colic is recognized as one of the most common causes of non-obstetric hospitalization during pregnancy. Furthermore, both symptomatic urolithiasis and its management have been associated with preterm labor, markedly amplifying the "typical" concerns of either pregnancy or stone passage alone. As those specializing in the care of stone disease are likely to be involved in the management of suspected renal colic in the pregnant woman, it is imperative to be informed of strategies and pitfalls in the evaluation and treatment of urinary tract stones in this population.

Compliance with American Urological Association Guidelines for Post-Percutaneous Nephrolithotomy Antibiotics Does Not Appear to Increase Rates of Infection.

PURPOSE: We compared infection rates after percutaneous nephrolithotomy in a group of patients without a history of infection or struvite calculi who received 24 hours or less of antibiotics postoperatively (ie compliance with AUA guidelines) vs a group that received 5 to 7 days of antibiotics postoperatively. MATERIALS AND METHODS: We retrospectively reviewed the records of consecutive percutaneous nephrolithotomy procedures in patients without a history of urinary tract infection. Group 1 received 24 hours or less of antibiotics postoperatively and group 2 received a mean of 6 days of antibiotics postoperatively. RESULTS: A total of 52 patients in group 1 (24 hours or less of antibiotics) and 30 in group 2 (mean 6 days of antibiotics) met study inclusion criteria. In 5 group 1 patients (9.6%) fever developed within 72 hours of percutaneous nephrolithotomy but none demonstrated bacteriuria or bacteremia on cultures. No patient in group 1 was treated for urinary tract infection on postoperative days 3 to 14. In 4 group 2 patients (13.3%) fever developed within 72 hours of percutaneous nephrolithotomy. A single patient showed bacteriuria (less than 10,000 cfu mixed gram-positive bacteria) on culture while no patient demonstrated bacteremia. No patient in group 2 was treated for urinary tract infection on postoperative days 3 to 14. There was no difference in stone-free rates or the need for additional procedures between the 2 groups. CONCLUSIONS: In this pilot series compliance with AUA guidelines for antibiotic prophylaxis did not result in higher rates of infection than in a comparable group of 30 patients who received approximately 6 days of antibiotics postoperatively.

Ndfip1 enforces a requirement for CD28 costimulation by limiting IL-2 production.

Although the pathways that permit IL-2 production and the full activation of T cells upon Ag encounter are fairly well defined, the negative regulatory circuits that limit these pathways are poorly understood. In this study, we show that the E3 ubiquitin ligase adaptor Ndfip1 directs one such negative regulatory circuit. T cells lacking Ndfip1 produce IL-2, upregulate IL-2Rα, and proliferate, in the absence of CD28 costimulation. Furthermore, T cells in mice lacking both Ndfip1 and CD28 become activated, produce IL-4, and drive inflammation at barrier surfaces. Ndfip1 constrains T cell activation by limiting the duration of IL-2 mRNA expression after TCR stimulation. Ndfip1 and IL-2 have a similar expression pattern, and, following TCR stimulation, expression of both Ndfip1 and IL-2 requires the activity of NFAT and Erk. Taken together, these data support a negative regulatory circuit in which factors that induce IL-2 expression downstream of TCR engagement also induce the expression of Ndfip1 to limit the extent of IL-2 production and, thus, dampen T cell activation.

Combination of error-prone PCR (epPCR) and Circular Polymerase Extension Cloning (CPEC) for improving the coverage of random mutagenesis libraries.

Random mutagenesis, such as error-prone PCR (epPCR), is a technique capable of generating a wide variety of a single gene. However, epPCR can produce a large number of mutated gene variants, posing a challenge in ligating these mutated PCR products into plasmid vectors. Typically, the primers for mutagenic PCRs incorporate artificial restriction enzyme sites compatible with chosen plasmids. Products are cleaved and ligated to linearized plasmids, then recircularized by DNA ligase. However, this cut-and-paste method known as ligation-dependent process cloning (LDCP), has limited efficiency, as the loss of potential mutants is inevitable leading to a significant reduction in the library's breadth. An alternative to LDCP is the circular polymerase extension cloning (CPEC) method. This technique involves a reaction where a high-fidelity DNA polymerase extends the overlapping regions between the insert and vector, forming a circular molecule. In this study, our objective was to compare the traditional cut-and-paste enzymatic method with CPEC in producing a variant library from the gene encoding the red fluorescent protein (DsRed2) obtained by epPCR. Our findings suggest that CPEC can accelerate the cloning process in gene library generation, enabling the acquisition of a greater number of gene variants compared to methods reliant on restriction enzymes.