Translating Senotherapeutic Interventions into the Clinic with Emerging Proteomic Technologies.

Cellular senescence is a state of irreversible growth arrest with profound phenotypic changes, including the senescence-associated secretory phenotype (SASP). Senescent cell accumulation contributes to aging and many pathologies including chronic inflammation, type 2 diabetes, cancer, and neurodegeneration. Targeted removal of senescent cells in preclinical models promotes health and longevity, suggesting that the selective elimination of senescent cells is a promising therapeutic approach for mitigating a myriad of age-related pathologies in humans. However, moving senescence-targeting drugs (senotherapeutics) into the clinic will require therapeutic targets and biomarkers, fueled by an improved understanding of the complex and dynamic biology of senescent cell populations and their molecular profiles, as well as the mechanisms underlying the emergence and maintenance of senescence cells and the SASP. Advances in mass spectrometry-based proteomic technologies and workflows have the potential to address these needs. Here, we review the state of translational senescence research and how proteomic approaches have added to our knowledge of senescence biology to date. Further, we lay out a roadmap from fundamental biological discovery to the clinical translation of senotherapeutic approaches through the development and application of emerging proteomic technologies, including targeted and untargeted proteomic approaches, bottom-up and top-down methods, stability proteomics, and surfaceomics. These technologies are integral for probing the cellular composition and dynamics of senescent cells and, ultimately, the development of senotype-specific biomarkers and senotherapeutics (senolytics and senomorphics). This review aims to highlight emerging areas and applications of proteomics that will aid in exploring new senescent cell biology and the future translation of senotherapeutics.

Effects of CYP3A4 and CYP2C9 genotype on systemic anastrozole and fulvestrant concentrations in SWOG S0226.

Objective & methods: This study tested associations of genotype-predicted activity of CYP3A4, other pharmacogenes, SLC28A7 (rs11648166) and ALPPL2 (rs28845026) with systemic concentrations of the endocrine therapies anastrozole and fulvestrant in SWOG S0226 trial participants. Results: Participants in the anastrozole-only arm with low CYP3A4 activity (i.e. CYP3A4*22 carriers) had higher systemic anastrozole concentrations than patients with high CYP3A4 activity (ß-coefficient = 10.03; 95% CI: 1.42, 18.6; p = 0.025). In an exploratory analysis, participants with low CYP2C9 activity had lower anastrozole concentrations and higher fulvestrant concentrations than participants with high CYP2C9 activity. Conclusion: Inherited genetic variation in CYP3A4 and CYP2C9 may affect concentrations of endocrine therapy and may be useful to personalize dosing and improve treatment outcomes.

Solid Renal Masses in Transplanted Allograft Kidneys: A Closer Look at the Epidemiology and Management.

The objective of this review is to explore the available literature on solid renal masses (SRMs) in transplant allograft kidneys to better understand the epidemiology and management of these tumors. A literature review using PubMed was performed according to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) methodology. Fifty-six relevant studies were identified from 1988 to 2015. A total of 174 SRMs in 163 patients were identified, with a mean tumor size of 2.75 cm (range 0.5-9.0 cm). Tumor histology was available for 164 (94.3%) tumors: clear cell renal cell carcinoma (RCC; 45.7%), papillary RCC (42.1%), chromophobe RCC (3%), and others (9.1%). Tumors were managed by partial nephrectomy (67.5%), radical nephrectomy (19.4%), percutaneous radiofrequency ablation (10.4%), and percutaneous cryoablation (2.4%). Of the 131 patients (80.3%) who underwent nephron-sparing interventions, 10 (7.6%) returned to dialysis and eight (6.1%) developed tumor recurrence over a mean follow-up of 2.85 years. Of the 110 patients (67.5%) who underwent partial nephrectomy, 3.6% developed a local recurrence during a mean follow-up of 3.12 years. The current management of SRMs in allograft kidneys mirrors management in the nontransplant population, with notable findings including an increased rate of papillary RCC and similar recurrence rates after partial nephrectomy in the transplant population despite complex surgical anatomy.

Pulmonary embolus originating from renal vein stump thrombus after laparoscopic living donor nephrectomy: case report.

BACKGROUND: Pulmonary embolism (PE) is an uncommon, life-threatening complication after living donor nephrectomy (LDN), and is considered among the most common causes for donor mortality. Most cases of postoperative PEs are thought to originate in deep venous thrombosis (DVT) of the lower extremities. CASE REPORT: A 56-year-old, healthy woman underwent laparoscopic left LDN. Her postoperative course was complicated by PE, presenting at postoperative day 7. Doppler ultrasonography of her lower extremities did not demonstrate DVT. Both transthoracic echocardiogram and contrast-enhanced computed tomography demonstrated a floating thrombus within the inferior vena cava (IVC) originating from a thrombus in the left renal vein stump. Symptoms resolved with systemic anticoagulation. Repeat transesophageal echocardiography demonstrated resolution of the IVC thrombus. CONCLUSIONS: Thrombus originating in left renal vein stump should be considered in patients who develop PE after LDN, especially when lower extremity DVT is not demonstrated.

Cryptococcal infection presenting as cellulitis in a renal transplant recipient.

Opportunistic infections of skin and soft tissue represent a rare but serious complication following solid organ transplantation. We report a case of severe soft tissue infection caused by Cryptococcus neoformans in a renal transplant recipient. Physicians need to consider the possibility of opportunistic pathogens when managing infections in immunocompromised hosts, especially when symptoms persist despite seemingly appropriate empiric antimicrobial therapy. Tissue sampling for histological and microbiological evaluation is usually necessary to establish a diagnosis.

Characterization of urinary peptide biomarkers of acute rejection in renal allografts.

We previously demonstrated that 4.7 kDa and 4.4 kDa peptides are useful in diagnosing acute rejection in renal transplant recipients. The aim of this study was to characterize these polypeptides and assess their potential as biomarkers. The polypeptides were identified as human beta-Defensin-1 (4.7 kDa) and alpha-1-antichymotrypsin (4.4 kDa), by tandem mass spectrometry and ProteinChip immunoassay. The urinary abundance of both polypeptides, assessed using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS), revealed a reduction in beta-Defensin-1 while alpha-1-antichymotrypsin increased in patients with rejection (p < 0.05) compared with clinically stable transplants. The area under the curve (AUC) for the receiver operator characteristic (ROC) curve for the diagnosis of rejection for the ratio of both peptides combined was 0.912. Longitudinal analysis confirmed a reduction in beta-Defensin-1 with a reciprocal increase in alpha-1-antichymotrypsin as rejection developed. The difference in urinary beta-Defensin-1 levels quantified by radioimmunoassay was 176.8 +/- 122.3 pg/mL in stable patients compared with 83.2 +/- 52.2 pg/mL in patients with acute rejection, with an ROC AUC of 0.749 (p < 0.01). Immunohistochemistry (IHC) confirmed reduced beta-Defensin-1 expression in the renal parenchyma of patients experiencing acute rejection. In conclusion, the ratio of beta-Defensin-1 and alpha-1-antichymotrypsin excretion in the urine is a novel, potentially useful candidate biomarkers of acute rejection.

East Ireland 1980-1994: epidemiology of neural tube defects.

STUDY OBJECTIVE: The objective of the study was to describe the epidemiology of neural tube defects (NTD) in the eastern region of Ireland using the EUROCAT register of congenital malformations. DESIGN, SETTING AND PATIENTS: EUROCAT registries monitor the prevalence of congenital anomalies in defined populations using multiple sources for case ascertainment. All cases of NTD on the Dublin EUROCAT register born between 1980 and 1994 were extracted and analysed. The crude birth prevalence rate for all NTD, spina bifida, anencephaly and encephalocoele were calculated for each year. Parameters measured were: sex ratio, stillbirth rate, proportion of low birth-weight babies (< 2500 g) and the proportion who were premature (< 37 weeks gestation). MAIN RESULTS: Of 821 NTD cases, 419 (51.0%) had spina bifida, 322 (39.2%) had anencephaly, 69 (8.4%) had encephalocoele and 11 (1.3%) were iniencephalic. The crude birth prevalence of NTD decreased fourfold from 46.9/10,000 births in 1980 to 11.6/10,000 in 1994. The downward trend ceased during the early 1990's. Younger mothers had significantly higher rates of NTD affected births. Twenty two per cent of NTD cases had additional non-central nervous system anomalies. In 40 cases, there was a previous family history of NTD in siblings. Seasonal effects in birth prevalence were observed. Birth notification was the most frequent mechanism of ascertainment. CONCLUSION: There was a marked fall in the birth prevalence of NTD during the 15 year period. This change was real and not accounted for by pre-natal screening and diagnostic practises with termination of pregnancy, which is not legally permissible in Ireland. Dietary factors may have had an influence. Rates of NTD in this region are still higher than many other parts of Europe. Primary prevention strategies through increased folic acid intake are necessary to further reduce NTD affected births.

Renal transplantation in multiple myeloma. Case report and review of the literature.

A patient with end-stage renal disease and known benign monoclonal gammopathy underwent kidney transplantation at Westchester County Medical Center, Valhalla, NY. After surgery, during routine follow-up, the patient had laboratory evidence of frank multiple myeloma. However, she did not show any clinical signs or symptoms of the disease. Four years later, the patient is asymptomatic and continues to have stable renal function. As a result of our experience, and that of others, we support transplantation as a viable option for patients with multiple myeloma.

Gitelman's syndrome (Bartter's variant) maps to the thiazide-sensitive cotransporter gene locus on chromosome 16q13 in a large kindred.

A defect in distal renal tubular sodium chloride handling is thought to be responsible for the clinical phenotype of Gitelman's syndrome, a variant of Bartter's syndrome. To study the possible involvement of the renal thiazide-sensitive NaCl cotransporter gene in the syndrome, a linkage analysis study in the largest reported kindred with the syndrome was performed. A human homolog of rat thiazide-sensitive cotransporter was cloned and mapped to chromosome 16q13 by fluorescent in situ hybridization. All 17 family members in two generations were genotyped at loci in this region. There were no recombinants observed between the Gitelman's syndrome phenotype and inheritance of D16S408 alleles, yielding a lod score of 3.88 at Q = 0. By contrast, recombinants were observed between Gitelman's syndrome and the flanking markers D16S419 and D16S400, localizing the responsible gene in this family to a 15 centimorgan region on chromosome 16q. These genetic data, together with current understanding of the molecular physiology of the thiazide-sensitive cotransporter, are strong evidence that the latter is defective in this kindred with Gitelman's syndrome.

Malacoplakia of the prostate in a renal transplant recipient. A complicated course.

Extrarenal malacoplakia in renal transplant recipients is generally associated with a good prognosis. We report the first case of malacoplakia of the prostate in a renal transplant recipient that was associated with a complicated course despite prophylactic antibiotic therapy and reduction of immunosuppression. Malacoplakia in a renal transplant recipient is not always benign.

Prevention of hypokalemia caused by diuretics.

Hypokalemia induced by the use of diuretics is common. Those at risk include the elderly, women, patients with edematous states, and patients in whom higher doses and/or the more potent agents are used. Prevention should include a low-salt diet rich in potassium, magnesium, and chloride (either through foods enriched with these elements or through potassium chloride supplements) and use of low doses of short-acting diuretics in the treatment of mild to moderate hypertension. The subgroup of hypertensive patients in whom hypokalemia develops despite these recommendations may benefit from a change to the potassium-sparing diuretic spironolactone or substitution of diuretics with alternative first-line drugs.

Deep vein thrombosis in end-stage renal disease.

Patients who have end-stage renal disease have an acquired platelet dysfunction that leads to prolonged bleeding time and that may put them at risk for bleeding. This platelet dysfunction is manifest in reduced platelet adhesion and impaired platelet aggregation. As a result, a bleeding tendency exists in end-stage renal disease, and understandably deep vein thrombosis is vanishingly rare in patients with this condition. We present three end-stage renal disease patients undergoing maintenance hemodialysis who developed deep vein thrombosis. These cases illustrate that in some patients with end-stage renal disease, the pro thrombotic forces may be so profound that they overwhelm the bleeding tendency.

The influence of delayed renal allograft function on long-term outcome in the cyclosporine era.

The influence of delayed renal graft function on long-term allograft outcome remains uncertain. All 495 cyclosporine treated cadaver donor renal transplants within a single center were analyzed with respect to dialysis dependence in the early posttransplant period. When compared with immediate allograft function, dialysis dependence for more than one week posttransplant was associated with prolonged cold ischemia time (27 +/- 11 vs 32 +/- 12 hours), cytotoxic antibodies > 30% (14% vs 25%), black race (29% vs 41%), increased incidence of acute rejection in the first year posttransplant (31% vs 67%) and inferior 1-year (85% vs 52%) and 5-year (68% vs 33%) graft survival among primary transplants. No adverse effect however was noted on renal function in long-term survivors.

Assessment of hypocalcemia and hypercalcemia.

Methodologic aspects including causes of factitious hypocalcemia and hypercalcemia are summarized. The differential diagnosis of hypocalcemia is reviewed under three main headings: hypoalbuminemia, hypocalcemia with decreased parathyroid hormone (PTH) action or activity, and hypocalcemia with normal PTH action and activity. The differential diagnosis of hypercalcemia is subdivided into three broad categories: hyperproteinemia, PTH-mediated hypercalcemia, and non-PTH-mediated hypercalcemia. The causes of hypocalcemia and hypercalcemia are outlined with a focus on pathophysiology and clinicochemical sequelae. A laboratory perspective is emphasized in outlining management strategies.