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Background After initiation of antiviral therapy (ART), plasma HIV-1 RNA is usually undetectable after one month. In rare cases, viral suppression may not be achieved despite good adherence and with virus susceptible to ART. We used ultra-deep Primer ID next gen sequencing (NGS) to study the viral population and evolution of HIV-1 in two patients with persistent viremia on intensive ART. Methods We extracted viral RNA from plasma samples collected at multiple timepoints over the duration of the treatment from two patients (VEX1 and VEX2). We constructed Primer ID NGS libraries covering part of the pol gene and the env V1/V3 region and sequenced them on the Illumina MiSeq platform. We used the ‘tcs’ pipeline to construct template consensus sequences (TCS) for each region, and searched for drug resistance mutations (DRMs). The Geno2pheno pipeline was used to predict co-receptor tropisms. Results Patient VEX1 was followed for two years on ART. VEX2 restarted ART in the hospital and received directly observed therapy for nearly 6 months. Both had over 1 million viral copies/mL at the initiation of ART, and the CD4 cell counts were extremely low. Their viral loads slowly declined to approx. 10,000 copies/mL at the end of follow-up but complete viral suppression was not achieved for either patient despite appropriate ART. DRMs were not detected in either patient throughout the treatment with detection sensitivity as low as 0.1%. The sequencing results for VEX1 showed that there were both X4- and R5-tropic viruses at all timepoints and R5 viruses decayed much more slowly than X4 viruses, up to 35-fold more slowly in the initial 3 months of ART. Phylogenetic analysis revealed that the persistent R5 viruses were largely clonal while little clonality was found in the persistent X4 virus. In VEX2, all viruses were R5-tropic. There were three major distinct lineages in the viral population, and two of them completely disappeared after the initiation of ART while the other lineage persisted throughout therapy (Fig 1). The persistent lineage in VEX2 was highly clonal. Pooled maximum likelihood tree at env V1V3 region of 3 time points from patient VEX2. Sequences in red were from 2018 when we obtained the initial specimen from the patient. Sequences in blue were from Sept 2021 when the patient restarted ART. Sequences in green were from Oct 2021 when the patient were on ART for 3 weeks. The patient had 3 major viral lineages, L1, L2, and L3. From 2018 to Sept 2021, we could see the viral evolution of the three lineages (blue arrows). After 3 weeks of ART, L1 and L2 disappeared, while the L3 persisted. We could also see that L3 had several clonal populations which did not respond to ART nor evolve from 2018 to 2021. The additional sequences from Nov 2021 to Feb 2022 were identical to the sequences obtained on Oct 2021 (not shown on this figure). Conclusion Our study shows that persistent viremia on ART can come from clonal viral lineages that carry no DRMs. Clonally expanded and infected host cells might contribute to the phenomenon. Further study is needed to explore the origin of these clonal viral genomes. Disclosures Jonathan Parr, MD, Abbott Laboratories: Donation of laboratory testing and reagents|Gilead Sciences: Grant/Research Support|Virology Education: Honoraria|World Health Organization: Advisor/Consultant|World Health Organization: Grant/Research Support David A. Wohl, M.D., Gilead: Advisor/Consultant|Gilead: Grant/Research Support|Janssen: Advisor/Consultant|Lilly: Grant/Research Support|Merck: Grant/Research Support|ViiV: Advisor/Consultant|ViiV: Grant/Research Support Joseph J. Eron, MD, Adagio Therapeutics: data safety monitoring committee|Gilead Sciences: Advisor/Consultant|Gilead Sciences: Grant/Research Support|Glaxo Smith Kline: Advisor/Consultant|Merck: Advisor/Consultant|ViiV Healthcare: Advisor/Consultant|ViiV Healthcare: Grant/Research Support.

2,4‐Thiazolidinedione (TZD) is a privileged and highly utilised scaffold for the development of pharmaceutically active compounds. This sulfur‐containing heterocycle is a versatile pharmacophore that confers a diverse range of pharmacological activities. TZD has been shown to exhibit biological action towards a vast range of targets interesting to medicinal chemists. In this review, we attempt to provide insight into both the historical conventional and the use of novel methodologies to synthesise the TZD core framework. Further to this, synthetic procedures utilised to substitute the TZD molecule at the activated methylene C5 and N3 position are reviewed. Finally, research into developing clinical agents, which act as modulators of peroxisome proliferator‐activated receptors gamma (PPARγ), protein tyrosine phosphatase 1B (PTP1B) and aldose reductase 2 (ALR2), are discussed. These are the three most targeted receptors for the treatment of diabetes mellitus (DM)., A key scaffold: 2,4‐Thiazolidinedione is a widely recognised structural motif present in many therapeutic agents used for the treatment of diabetes mellitus. This review summarises some of the methods used to prepare biologically active members of this class and discusses the pharmacological potency of these compounds.

Background Total shoulder arthroplasty is an accepted treatment for glenohumeral osteoarthritis. The Arthrex Eclipse shoulder prosthesis is a stemless, canal-sparing humeral prosthesis with bone ingrowth capacity on the trunnion, as well as through the fenestrated hollow screw, that provides both diaphyseal and metaphyseal load sharing and fixation. Methods Between 2013 and 2018, 16 sites in the United States enrolled 327 patients (Eclipse in 237 and Arthrex Univers II in 90). All patients had glenohumeral arthritis refractory to nonsurgical care. Strict exclusion criteria were applied to avoid confounding factors such as severe patient comorbidities, arthritis not consistent with osteoarthritis, and medical or prior surgical treatments that may have affected outcomes. Patients were randomized to the Eclipse or Univers II group via block randomization. Results In total, 149 Eclipse and 76 Univers II patients reached 2-year follow-up (139 Eclipse patients [93.3%] and 68 Univers II patients [89.5%] had complete data). The success rate using the Composite Clinical Success score was 95% in the Eclipse group vs. 89.7% in the Univers II group. No patient exhibited radiographic evidence of substantial humeral radiolucency, humeral migration, or subsidence at any point. Reoperations were performed in 7 patients (3.2%) in the Eclipse group and 3 (3.8%) in the Univers II group. Conclusion The Arthrex Eclipse shoulder prosthesis is a safe and effective humeral implant for patients with glenohumeral arthritis at 2-year follow-up, with no differences in outcomes compared with the Univers II shoulder prosthesis.

In the female rat, sexual receptivity (lordosis) can be facilitated by sequential activation of estrogen receptor (ER) α and G protein-coupled estrogen receptor 1 (GPER) by estradiol. In the estradiol benzoate (EB) primed ovariectomized (OVX) rat, EB initially binds to ERα in the plasma membrane that complexes with and transactivates metabotropic glutamate receptor 1a to activate β-endorphin neurons in the arcuate nucleus of the hypothalamus (ARH) that project to the medial preoptic nucleus (MPN). This activates MPN μ-opioid receptors (MOP), inhibiting lordosis. Infusion of non-esterified 17β-estradiol into the ARH rapidly reduces MPN MOP activation and facilitates lordosis via GPER. Tamoxifen (TAM) and ICI 182,780 (ICI) are selective estrogen receptor modulators that activate GPER. Therefore, we tested the hypothesis that TAM and ICI rapidly facilitate lordosis via activation of GPER in the ARH. Our first experiment demonstrated that injection of TAM intraperitoneal, or ICI into the lateral ventricle, deactivated MPN MOP and facilitated lordosis in EB-primed rats. We then tested whether TAM and ICI were acting rapidly through a GPER dependent pathway in the ARH. In EB-primed rats, ARH infusion of either TAM or ICI facilitated lordosis and reduced MPN MOP activation within 30 min compared to controls. These effects were blocked by pretreatment with the GPER antagonist, G15. Our findings demonstrate that TAM and ICI deactivate MPN MOP and facilitate lordosis in a GPER dependent manner. Thus, TAM and ICI may activate GPER in the CNS to produce estrogenic actions in neural circuits that modulate physiology and behavior.

Pensions automatic enrolment is 5 years old. While the rollout is complete, employers duties are ongoing. With stiff penalties for non-compliance, care homes need to understand their obligations towards their employees' pensions

The objective was to determine the effects of immunomodulatory feed ingredient during post-weaning on cortisol concentration and fecal microbial populations of beef heifers. Commercial Angus heifers (n = 72) from two AI sires were blocked (n = 9) by sire and BW, randomly assigned to one of two pens (4 heifers/pen) per block, then assigned to treatments. Heifers were fed twice daily from d 0 to 60 (gain 0.75 kg/day) and topdressed once daily with either 72g of Celmanax (CEL) or corn germ (CON; corn germ meal) per pen. After 60 days, two heifers per pen (n = 32) were randomly selected for a transportation challenge. Fecal grab samples were collected on d 0 and 69 of treatment, hr -24, 0, 24 of the challenge and 7 d post-challenge. Serum samples were collected at h 0, 4, 8, and 12 of the challenge. Clostridia and E. coli were enumerated from fecal samples using selective media. Isolates (≤ five isolates from each media per sample) were genetically tested to determine if they were C. perfringens or pathogenic E. coli. Fecal samples were enriched for detection of Salmonella. Pen was the experimental unit and data was analyzed by ANOVA or repeated measures analysis. Following treatment, decreased (P ≤ 0.05) populations of total E. coli, Salmonella, and C. perfringens were observed in CEL heifers compared to CON heifers, whereas clostridia and pathogenic E. coli were not different (P > 0.05) between treatments. Transportation stress increased (P ≤ 0.05) populations of clostridia, C. perfringens, total E. coli, and Salmonella, but decreased (P = 0.0252) pathogenic E. coli counts. Cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to CON heifers throughout the challenge. In summary, supplementation of Celmanax post-weaning altered microbial populations and cortisol concentrations were reduced during transportation in beef heifers

Workplace pensions have changed radically over the last few years, and will continue to do so until at least February 2018. In this article, Nathan Long explains the obligations that employers are under, and how these are likely to change over the next few years

In female rats sexual receptivity (lordosis) can be induced with either a single large dose of estradiol benzoate (EB), or a priming dose of EB that does not induce sexual receptivity followed by 17β-estradiol (E2). Estradiol priming initially inhibits lordosis through a multi-synaptic circuit originating in the arcuate nucleus of the hypothalamus (ARH) that activates and internalizes μ-opioid receptors (MOR) in medial preoptic nucleus (MPN) neurons. Lordosis is facilitated when MPN MOR are deactivated after the initial estradiol-induced activation. We tested the hypothesis that E2 given 47.5 hours post EB acts rapidly through G protein-coupled estrogen receptor 1 (GPER) in the ARH to deactivate MPN MOR and facilitate lordosis. Ovariectomized Long Evans rats implanted with a third ventricle cannula were primed with 2 μg EB. DMSO control, E2, or G1 (GPER selective agonist) was infused 47.5 hours later, and rats were tested for sexual receptivity. E2 and G1 infusions significantly increased levels of sexual receptivity compared to DMSO controls and pretreatment with G15 (GPER antagonist) blocked the facilitation of sexual receptivity. Brains were processed for MPN MOR immunohistochemistry to measure MPN MOR activation levels. E2 and G1 both significantly reduced MPN MOR activation compared to DMSO controls, while pretreatment with G15 blocked MPN MOR deactivation. In another group of EB treated ovariectomized rats, GPER immunofluorescence positive staining was observed throughout the ARH. Together these data indicate that in the 2 μg EB primed rat, E2 rapidly signals through GPER in the ARH to deactivate MPN MOR and facilitate lordosis.

This paper describes the preparation method as well as the in vitro and in vivo evaluation of a novel flexible glucose biosensor designed for long-term subcutaneous implantation. An epoxy-enhanced polyurethane membrane, which includes ca. 30-40% epoxy resin adhesive and 50-70% polyurethane, has been developed and used for the first time as the outer protective membrane of the sensor. This new membrane was developed to increase the in vivo durability and lifetime of implantable biosensors. This epoxy-polyurethane membrane was shown to be porous and is of excellent durability. A sensor with such a membrane shows excellent long-term stability and can last for 4-8 months in solutions at room temperature. To verify the in vivo performance of the sensor, nine sensors were implanted in three rats and tested regularly. Eight sensors kept functioning well in the rats for 10-56 days. The ninth sensor was damaged during implantation. All original sensitivity data as well as four response curves obtained at days 7, 17, 52 and 56, respectively are presented.

Transcutaneous and embedded devices were developed for use in characterizing the in vivo performance of subcutaneously implanted glucose sensors. The devices were used as a portal for accessing electrochemical glucose sensors from the exterior. They were designed to prevent the sensors from being pulled out of the animals and the sensor leads from breaking. Development of the devices took into consideration rodent mobility, infection control, and animal welfare balanced with sensor durability, accessibility, and functionality.Our approach was developed over five animal protocols spanning a period of 6 months. A total of 68 sensors were implanted with 60 associated devices in 22 Sprague-Dawley outbred rats.The average sensor lifetime was 11.2 +/- 3.1 days with a maximum of 56 days. All-cause sensor failure averaged one sensor per day. As implantation devices were modified, failure attributable to the device was decreased by 40%. The resulting devices showed good durability and allowed for easy sensor access and testing.These data represent baseline sensor function against which future sensor improvements may be measured. The new devices and techniques described should be a valuable tool in the development of continuous glucose sensors.

The clinician must be the ultimate medical detective when dealing with chronic optic neuropathies. History taking is crucial. Clinical examination may require supplementation with visual field testing, fluorescein angiography, ocular and orbital ultrasound imaging, CT and MR imaging, blood test data, and cerebrospinal fluid or tissue biopsy data to determine the specific diagnosis. This supplementation is labor-intensive and time-consuming; the visual loss usually will progress throughout the process, frustrating and frightening the patient and physician. The final common pathway is gradual optic atrophy; the appearance of the optic nerve is rarely adequate to determine the cause of the visual loss. This article includes tables that review diagnostic aids and therapies, and lists the frequency with which several disease entities were encountered over 15 years in one tertiary care neuro-ophthalmic practice. If a specific cause is discernible, then a specific therapy may be available. This approach has the best chance of saving the patient‘s vision with the least toxicity caused by erroneous trials. By necessity, the work-up for these patients is expensive, but the cost of not pursuing the cause is irrevocable, permanent blindness.

The relationship between tissue reactions to a subcutaneously implanted glucose sensor and the function of the sensor was evaluated over a period of 4-weeks using tubular, porous polyvinyl alcohol (PVA) sponges implanted subcutaneously in rats. The PVA sponges were used as scaffolds in which the foreign body response could develop. Coil-type glucose sensors were then placed in the center of the PVA sponges and tested on day 3, and weekly thereafter. In the first approach, the sensors were placed in the sponges still implanted in the rats and tested. In vivo glucose sensor sensitivity peaked at day 7 and steadily decreased until day 35. In the second approach, the sensors were placed in the explanted sponges and then tested. This test showed no sensor function after day 7, indicating that functional blood vessels are critical in maintaining any function whatsoever. In both cases the sensors themselves were never implanted to eliminate any potential in vivo degradation of the sensors that could have affected the outcome of this study. Sensors were then tested in absence of sponges and found to be working properly with no change from preimplantation sensitivity. Once sensor testing was concluded, the PVA sponge/tissue samples were prepared for quantitative histological analysis. It was determined that the increase in collagen deposition within the sponge correlated with the decrease in sensor sensitivity. It was also observed that natural angiogenesis (peak at day 14) did not overcome the barrier to glucose diffusion created by the fibrous capsule. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2008