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expressed in post-meiotic stages, including spermatids and spermatozoa in the testes, and the principal piece of the flagellum and acrosome in ejaculated spermatozoa. A case-control study revealed that 4.5% of infertile patients carry the G75C polymorphism, which causes an amino acid substitution in the encoded protein. Prevalence of PGAM4 SNPs in infertile and proven-fertile human populations were indicated in Table. Furthermore, an assay for enzymatic activity demonstrated that this polymorphism decreases the enzyme’s activity both in vitro and in vivo. CONCLUSIONS: These results suggest that PGAM4, an Xlinked retrogene, is a fundamental gene in human male reproduction and may escape meiotic sex chromosome inactivation. These findings provide fresh insight into elucidating the mechanisms of male infertility.

Cardiovascular disease is a major cause of morbidity and mortality in the USA. Traditional risk factors such as obesity, physical inactivity, and diet are used to screen for cardiovascular disease. However, these risk factors miss a significant population who are at risk for future cardiac events. Erectile dysfunction (ED) has many associated conditions in common with cardiovascular disease and has been shown to be an independent risk factor for cardiovascular. Measurements made on penile Doppler ultrasound (PDU), such as cavernosal artery peak systolic velocity (PSV), cavernosal artery intima-medial thickness, and the finding of cavernosal artery calcification, are indicators of generalized vascular disease. Thus, elements of PDU can identify men at higher risk for cardiovascular disease. This review outlines the proper technique for PDU and the literature supporting the use of PDU to predict cardiovascular disease in men with erectile dysfunction.

A new poly(ethylene glycol) derivative, 3-[methoxypoly(oxyethylene)]methylene furan, I, was prepared from the reaction of 3-furanmethanol with the mesylate of methoxypoly(oxyethylene) in tetrahydrofuran. The degree of end-group conversion, as determined by NMR spectroscopy, was 100%. The Diels-Alder reactions of I with N-phenylmaleimide, N-glycinylmaleimide, maleic anhydride, N,N'-hexamethylene bismaleimide, and diethyl acetylenedicarboxylate resulted in the corresponding adducts. For the adduct derived from I and N-phenylmaleimide, its thermal reversible character was confirmed by applying a retro-Diels-Alder reaction in the presence of a large excess of 2-methylfuran, which restored the initial polymer I quantitatively. The adduct obtained from I and N-glycinylmaleimide was converted into its succinimidyl ester and its hydrolysis rate in phosphate buffer (pH = 8) was determined. The reactivity of the adduct derived from I and N,N'-hexamethylene bismaleimide with benzyl mercaptan was also investigated.

Mucopolysaccharidosis I (MPS I) due to deficient alpha-L-iduronidase (IDUA) activity results in accumulation of glycosaminoglycans in many cells. Gene therapy could program liver to secrete enzyme with mannose 6-phosphate (M6P), and enzyme in blood could be taken up by other cells via the M6P receptor. Newborn MPS I mice were injected with 10(9) (high dose) or 10(8) (low dose) transducing units/kg of a retroviral vector (RV) expressing canine IDUA. Most animals achieved stable expression of IDUA in serum at 1240 +/- 147 and 110 +/- 31 units/ml, respectively. At 8 months, untreated MPS I mice had aortic insufficiency, increased bone mineral density (BMD), and reduced responses to sound and light. In contrast, MPS I mice that received high-dose RV had normal echocardiograms, BMD, auditory-evoked brain-stem responses, and electroretinograms. This is the first report of complete correction of these clinical manifestations in any model of mucopolysaccharidosis. Biochemical and pathologic evaluation confirmed that storage was reduced in these organs. Mice that received low-dose RV and achieved 30 units/ml of serum IDUA activity had no or only partial improvement. We conclude that high-dose neonatal gene therapy with an RV reduces some major clinical manifestations of MPS I in mice, but low dose is less effective.

Diels–Alder reactions of 5-[methoxypoly(oxyethylene)]-(3E)-1,3-pentadiene (1a) with maleic anhydride, diethyl acetylenedicarboxylate (DADC), and acrolein were investigated for the synthesis of new poly(ethylene glycol) derivatives. To facilitate the characterization of the derivatives, Diels–Alder reactions of 5-methoxyethoxy-(3E)-1,3-pentadiene (1b) with the aforementioned dienophiles were also studied. The reaction of o-toluidine with the cycloaddition product from maleic anhydride and 1b resulted in the corresponding amide products. The reactions of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone with cycloadducts derived from 1a and 1b with DADC resulted in the aromatization of the corresponding products. An NMR analysis of the adducts obtained from 1a and acrolein in water and from 1b and acrolein in water/acetonitrile (4:1 v/v) indicated a mixture of endo and exo, with the endo concentration being approximately 80%. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 1895–1902, 2005

Protein physical and chemical properties can be altered by polymer interaction. The presence of several high affinity binding sites on human serum albumin (HSA) makes it a possible target for many organic and polymer molecules. This study was designed to examine the interaction of HSA with poly(ethylene glycol) (PEG) in aqueous solution at physiological conditions. Fourier transform infrared, ultraviolet-visible, and CD spectroscopic methods were used to determine the polymer binding mode, the binding constant, and the effects of polymer complexation on protein secondary structure. The spectroscopic results showed that PEG is located along the polypeptide chains through H-bonding interactions with an overall affinity constant of K = 4.12 x 10(5) M(-1). The protein secondary structure showed no alterations at low PEG concentration (0.1 mM), whereas at high polymer content (1 mM), a reduction of alpha-helix from 59 (free HSA) to 53% and an increase of beta-turn from 11 (free HSA) to 22% occurred in the PEG-HSA complexes (infrared data). The CDSSTR program (CD data) also showed no major alterations of the protein secondary structure at low PEG concentrations (0.1 and 0.5 mM), while at high polymer content (1 mM), a major reduction of alpha-helix from 69 (free HSA) to 58% and an increase of beta-turn from 7 (free HSA) to 18% was observed.

A new method to synthesize vinyl ethers of methoxyoligooxyethylene, I, and methoxypoly(oxyethylene), II, is described. In this work, these derivatives were prepared by reacting the alkoxide of ethylene glycol vinyl ether with the tosylate (or mesylate) of oligo-/or methoxypoly(oxyethylene) in tetrahydrofuran.

Study Type – Therapy (case series) Level of Evidence 4 OBJECTIVE • To determine the pressure-flow characteristics of neobladders created in various configurations that may be constructed intra-abdominally. Complete intracorporeal neobladder construction has been previously described but is limited due to excessive operative time and the need for an advanced laparoscopic skill set. MATERIALS AND METHODS • Four neobladder configurations were constructed, each using 20 cm of human cadaveric small intestine. The standard hand sewn Studer pouch was compared with a circular loop, W-pouch, and U-pouch with stapled anastamoses. • Pressure flow studies were completed using the Aquarius TT UDS system (Laborie Medical Technologies, Toronto, Ontario) and each neobladder was filled to a pressure of 50 cm H2O. Neobladder change in pressure, capacity, and overall compliance were determined. RESULTS • The cystometric capacities of the stapled U-pouch, W-pouch, Circle pouch, and Studer pouch were 167.3 mL, 177.5 mL, 114 mL, and 145.2 mL respectively. The first increase in intravesical pressure was at 90.3 mL, 103 mL, 50 mL, and 85 mL. • The greatest compliance of 3.81 mL/cmH2O was demonstrated in the U-pouch, with the W-pouch revealing a compliance of 3.44 mL/cmH2O. • The least compliant neobladder was the circle pouch (2.24 mL/cmH20) followed by the standard Studer pouch (2.94 mL/cmH2O). CONCLUSION • The construction of an orthotopic neobladder must not only be technically feasible but maintain adequate capacity and compliance for optimal functioning. Pressure-flow studies demonstrated equivalent results in alternate neobladder configurations. Additional data is needed to determine feasibility in vivo.

Objectives Guidelines recommend hepatitis C virus (HCV) screening for all people living with HIV (PLWH). Understanding HCV testing practices may improve compliance with guidelines and can help identify areas for future intervention. Methods We evaluated HCV screening and unnecessary repeat HCV testing in 8,590 PLWH initiating care at 12 U.S. HIV clinics between 2006 and 2010, with follow-up through 2011. Multivariable logistic regression examined the association between patient factors and the outcomes: HCV screening (≥1 HCV antibody tests during the study period) and unnecessary repeat HCV testing (≥1 HCV antibody tests in patients with a prior positive test result). Results Overall, 82% of patients were screened for HCV, 18% of those screened were HCV antibody-positive, and 40% of HCV antibody-positive patients had unnecessary repeat HCV testing. The likelihood of being screened for HCV increased as the number of outpatient visits rose (adjusted odds ratio 1.02, 95% confidence interval 1.01–1.03). Compared to men who have sex with men (MSM), patients with injection drug use (IDU) were less likely to be screened for HCV (0.63, 0.52–0.78); while individuals with Medicaid were more likely to be screened than those with private insurance (1.30, 1.04–1.62). Patients with heterosexual (1.78, 1.20–2.65) and IDU (1.58, 1.06–2.34) risk compared to MSM, and those with higher numbers of outpatient (1.03, 1.01–1.04) and inpatient (1.09, 1.01–1.19) visits were at greatest risk of unnecessary HCV testing. Conclusions Additional efforts to improve compliance with HCV testing guidelines are needed. Leveraging health information technology may increase HCV screening and reduce unnecessary testing.

sarcoidosis, CA-125 antigen Sarcoidosis is a multisystem disease that is characterized by noncaseating epithelioid granulomas. Extrapulmonary sarcoidosis in addition to pulmonary involvement has been well described and occurs in 50% of patients. However, sarcoidosis without any pulmonary involvement is uncommon[1]. Cancer Antigen 125 (CA-125) is a tumor marker that is associated with ovarian malignancy. Sarcoidosis with elevated serum CA-125 has only been reported five times in the literature[2]. When sarcoidosis occurs in the absence of pulmonary involvement with elevated serum CA-125, it may mimic the presentation of disseminated ovarian carcinoma. This abdominal CT scan (Fig. 1) is from a 49-year-old, African American female with no known medical comorbidities, who presented with subjective chills and 20-lb weight loss in 6 months. She was found to have hepatomegaly, but no jaundice or stigmata of chronic liver

We have investigated the reaction chemistry of methoxypoly(oxyethylene)propionitrile, I, to synthesize methoxypoly(oxyethylene)propionic acid, II. We have found that II can be prepared by converting I first to the corresponding amide. Subsequent hydrolysis of the amide then yields II.

Despite the clinical success of PD-1 based therapies in human melanoma patients, the majority of patients do not have durable clinical benefit from anti-PD-1 monotherapy. A major challenge remains identifying which patients will respond to anti-PD-1 therapy and defining the underlying reasons for successful response versus treatment failure. Pre-existing T cell infiltration and/or PD-L1 expression in tumors may predict clinical responses; however, the use of blood-based profiling to understand the immunologic mechanism of PD-1 blockade has been less explored. Here we used detailed immune profiling of peripheral blood from stage IV melanoma patients before and after pembrolizumab (pembro), and identified pharmacodynamic changes in circulating exhausted-phenotype CD8 T cells (TEX). Robust induction of Ki67 in this subset of circulating CD8 T cells post-therapy (re-invigoration) occurred in 78% of patients indicating strong, on target immunological effects of PD-1 blockade in most patients studied here. Despite this high immunological response rate, the objective clinical response rate in this cohort was less than 40%. Ki67 in CD8 T cells alone did not predict clinical outcomes and, in fact, higher systemic immune activation at baseline was associated with lower overall survival. Rather, the magnitude of re-invigoration of circulating TEX in relation to pre-treatment tumor burden correlated with clinical response. We identified a TEX re-invigoration to tumor burden ratio which could be used to predict clinical response and overall survival as early as 6 weeks post therapy. Consistent observations were found in a second independent cohort and suggest that clinical failure of PD-1 blockade in many patients may not solely be due to an inability to induce immune re-invigoration but rather, an imbalance between T cell re-invigoration and tumor burden. Thus, by focused profiling of a mechanistically relevant circulating T cell subpopulation calibrated to pre-treatment disease burden, we identify a clinically accessible predictor of response to PD-1 blockade. These findings also provide a framework for dissecting distinct types of treatment failures in melanoma and have implications for stratifying patients into additional immunotherapeutic treatment approaches. Citation Format: Alexander Huang, Michael A. Postow, Robert J. Orlowski, Rosemarie Mick, Bertram Bengsch, Sasi Manne, Wei Xu, Shannon Harmon, Matthew Adamow, Deborah Kuk, Katherine Panangeas, Cristina Carerra, Phillip Wong, Felix Quagliarello, Kristen E. Pauken, Ramin S. Herati, Suzanne McGettigan, Shawn Kothari, Sangeeth M. George, Brandon Wenz, Kurt D'Andrea, Xiaowei Xu, Ravi K. Amaravadi, Giorgos Karakousis, Lynn M. Schuchter, Katherine L. Nathanson, Jedd D. Wolchok, Tara C. Gangadhar, John Wherry. Peripheral blood immune profiling of anti-PD-1 therapy in human melanoma reveals a link between T cell re-invigoration and tumor burden that predicts response [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr PR05.

T follicular helper cells provide essential B cell help within the germinal center (GC), and these cells have been shown to populate the follicular mantel once the GC reaction is complete. The significance and ontology of circulating Tfh (cTfh) is less clear. Studies in humans have suggested subsets of cTfh that correlate with humoral responses in chronic infection (HIV, malaria) and in response to acute challenge (influenza vaccination). We have previously described a subset of CXCR5+PD1+ cTfh that expresses both ICOS and CD38. This population was significantly increased 7 days after influenza vaccination as well as in patients infected with HIV. Single cell transcriptional analysis of the ICOS+CD38+ cTfh after influenza vaccination demonstrated more frequent expression of the GC Tfh transcripts CMAF, BCL6, and CD200 than was found in the ICOS−CD38− cTfh, and bulk transcriptional analysis indicated higher expression of IL-21. These data suggested that ICOS and CD38 mark the rare population of cTfh that are more recent émigrés from the lymph nodes. To evaluate this possibility, we analyzed Tfh in an immune compartment that is more proximal to the lymph nodes than peripheral blood: thoracic duct lymph (TDL). As expected, TDL Tfh had a higher frequency of ICOS+CD38+ cells than cTfh in matched peripheral blood. In addition, TDL had a population of CXCR5+PD1bright Tfh, previously only identified in lymphoid tissues. CXCR5+PD1bright Tfh in the TDL had a significantly increased frequency of ICOS+CD38+ cells. These data provide a critical missing link in the understanding of cTfh and suggest that the ICOS+CD38+ cTfh subset warrants further exploration as peripheral surrogates of the active immune responses within secondary lymphoid tissues.