Progesterone receptor membrane component-1 (PGRMC1) is the mediator of progesterone's antiapoptotic action in spontaneously immortalized granulosa cells as revealed by PGRMC1 small interfering ribonucleic acid treatment and functional analysis of PGRMC1 mutations

Progesterone (P4) receptor membrane component-1 (PGRMC1) and its binding partner, plasminogen activator inhibitor 1 RNA binding protein (PAIRBP1) are thought to form a complex that functions as membrane receptor for P4. The present investigations confirm PGRMC1's role in this membrane receptor complex by demonstrating that depleting PGMRC1 with PGRMC1 small interfering RNA results in a 60% decline in [(3)H]P4 binding and the loss of P4's antiapoptotic action. Studies conducted on partially purified GFP-PGRMC1 fusion protein indicate that [(3)H]P4 specifically binds to PGRMC1 at a single site with an apparent K(d) of about 35 nm. In addition, experiments using various deletion mutations reveal that the entire PGRMC1 molecule is required for maximal [(3)H]P4 binding and P4 responsiveness. Analysis of the binding data also suggests that the P4 binding site is within a segment of PGRMC1 that is composed of the transmembrane domain and the initial segment of the C terminus. Interestingly, PAIRBP1 appears to bind to the C terminus between amino acids 70-130, which is distal to the putative P4 binding site. Taken together, these data provide compelling evidence that PGRMC1 is the P4 binding protein that mediates P4's antiapoptotic action. Moreover, the deletion mutation studies indicate that each domain of PGRMC1 plays an essential role in modulating PGRMC1's capacity to both bind and respond to P4. Additional studies are required to more precisely delineate the role of each PGRMC1 domain in transducing P4's antiapoptotic action.     

Yield of Repeat 3D Angiography in Patients with Aneurysmal-Type Subarachnoid Hemorrhage

BACKGROUND AND PURPOSE:Aneurysmal-type subarachnoid hemorrhage is a serious disease with high morbidity and mortality. When no aneurysm is found, the patient remains at risk for rebleeding. Negative findings for SAH on angiography range from 2% to 24%. Most previous studies were based on conventional 2D imaging. 3D rotational angiography depicts more aneurysms than 2D angiography. The purpose of this study was to evaluate the yield of repeat 3D rotational angiography in patients with aneurysmal-type SAH with negative initial 3D rotational angiography findings and to classify the initial occult aneurysms.MATERIALS AND METHODS:Between March 2013 and January 2016, 292 patients with SAH and an aneurysmal bleeding pattern were admitted. Of these 292 patients, 30 (10.3%; 95% CI, 7.3%–14.3%) had initial negative 3D rotational angiography findings within 24 hours. These patients underwent a second 3D rotational angiography after 7–10 days.RESULTS:In 8 of 30 patients (26.7%; 95% CI, 14.0%–44.7%) with initial negative 3D rotational angiography findings, a ruptured aneurysm was found on repeat 3D rotational angiography. Three of 8 initial occult aneurysms were very small (1–2 mm), 2 were supraclinoid carotid artery dissecting aneurysms (2 and 8 mm), 2 were small (1 and 3 mm) basilar perforator aneurysms, and 1 was a 3-mm vertebral artery dissecting aneurysm.CONCLUSIONS:In 10% of patients with aneurysmal-type SAH, initial 3D rotational angiography findings were negative, and in 1 in 4, repeat 3D rotational angiography demonstrated a ruptured aneurysm. Initial occult aneurysms were dissecting aneurysms of perforators or main arteries or were very small (1–2 mm) or both. Our results indicate that repeat 3D rotational angiography is mandatory in patients with initial 3D rotational angiography findings negative for aneurysmal-type SAH.

Subarachnoid hemorrhage with an aneurysmal bleeding pattern (aSAH) is a serious disease with high morbidity and mortality. In 80%–90% of patients with aSAH, an aneurysm can be found as the cause of hemorrhage,¹ and early repair is advocated to prevent recurrent hemorrhage.The incidence of angiograms negative for aSAH has been reported to range from 2% to 24% in various studies.^2,3 In some of these patients, the source of the hemorrhage is an occult aneurysm, but intracranial artery dissections, dural arteriovenous malformations, micro-AVMs, trauma, bleeding disorders, substance abuse, or other causes should also be considered. Even though no aneurysm is found, these patients are at risk for early rebleed with an inherent risk of morbidity and death.⁴ An aneurysm is not depicted on first angiography for several reasons: very small aneurysms, small aneurysms with an intraluminal thrombus, aneurysms on arterial dissections, suboptimal image quality due to technical reasons or in uncooperative patients, and a missed diagnosis of an aneurysm.Repeat angiography is advocated to avoid missing a treatable cause of aSAH. Previous studies have demonstrated that repeat angiography after 7–10 days can depict an aneurysm in up to one-third of patients with initial negative angiography findings.^5–8 3D rotational angiography (3DRA) has been proved to depict more aneurysms than 2D DSA.^9,10 In our institution, diagnostic work-up of patients with aSAH now consists of 3DRA of all vessels within 24 hours. With negative findings, 3DRA is repeated. In this study, we evaluated the yield of repeat 3DRA in patients with aSAH with negative initial 3DRA findings. In addition, we classified the initial occult aneurysms.     

Effect of betulinic acid administration on TLR-9/NF-κB /IL-18 levels in experimental liver injury

Background/aim Acetaminophen (APAP), used in the composition of thousands of preparations, is the most commonly used analgesic and antipyretic drug. The present study aimed to investigate the potential protective effects of the betulinic acid (BA) treatment through an APAP-induced hepatotoxicity rat model, using inflammatory, biochemical, and histopathological parameters. Materials and methodsThe study consisted of four groups: control group, APAP group, BA group, and APAP+BA group. Experimental studies continued for fifteen days. Serum samples were analysed for glucose, total cholesterol (TChol), triglyceride (TG), low density lipoprotein (LDL), high density lipoprotein (HDL), aspartate amino transferase (AST), malondialdehyde (MDA), toll-like receptor-9 (TLR-9), nuclear factor kappa B (NF-κB), and interleukin-18 (IL-18).ResultsTLR9, IL-18, NF-κB, and MDA levels increased significantly in liver injury groups. These increases considerably decreased by the BA treatment. All groups showed immunopositivity for 8-hydroxy-2’–deoxyguanosine (8-OHdG) and interleukin (IL-1β) in the hepatocytes, inflammatory cells, and epithelial cells of bile ducts. Conclusion BA can be used as an effective agent in the prevention and treatment of acute liver diseases due to its inhibitory properties in multiple pathways and its potent antioxidant effects.     

Assessing joint involvement in haemophilia by clinical rheumatologic scores. A pilot study on similarities with subjects with psoriatic arthritis

Because of joint haemorrhages, severe haemophilia subjects often have limitations in their daily activities. Current orthopaedic scores (OJS) in haemophilia miss mild joint impairments and only pick up severe alterations. Twelve young severe haemophiliacs (20.25 ± 1.9 years of age), were evaluated for OJS as well as for indices employed in rheumatoid arthritis [28-joint Disease Activity Score (DAS-28), Ritchie index, Health Assessment Questionnaire (HAQ), visual analogue scale (VAS)], spondyloarthritis [Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), HAQ, VAS] and osteoarthritis [Knee injury and Osteoarthritis Outcome Score (KOOS), VAS]. Twenty-four matched apparently healthy subjects and 29 subjects with psoriatic arthritis (PsA) with oligoarticular involvement (one to three swollen joints) served as controls. In addition to the impairment of target joints (elbow, in five of five in those on-demand treatment; three, elbows; four, knee in those on secondary prophylaxis), HAQ (mean 0.71 ± 0.95) and VAS (3.12 ± 2.36) documented a quality of life and a perception of pain in haemophiliacs similar to that of PsA subjects (p = 0.061 and p = 0.063, respectively). Their Ritchie index did not differ from that of subjects with psoriatic arthritis (5.75 ± 8.1 vs 7.73 ± 9.22; p = 0.408), nor did the BASDAI score with respect to psoriatic arthritis patients (p = 0.105). Six of 12 haemophiliacs (50%) had KOOS values from 70 to 50 (significant function joint impairment); 3 of 12 (25%) showed DAS-28 values >3.2 (moderate disease activity), 2 of 12 (16.6%) severe disease activity (>5.1). All these indices significantly correlated with VAS and HAQ in haemophilia subjects. A rheumatologic assessment may help identify early polyarticular disease and subclinical abnormalities involving joints not usually studied (not target joint) in haemophiliacs. These pilot data provide the rationale for testing a systemic involvement in haemophiliacs by means of high-tech imaging and to start early-onset prophylaxis/treatment in this setting.     

Quantifying the risks and benefits of efavirenz use in HIV‐infected women of childbearing age in the USA

Objectives

The aim of the study was to quantify the benefits (life expectancy gains) and risks (efavirenz‐related teratogenicity) associated with using efavirenz in HIV‐infected women of childbearing age in the USA.

Methods

We used data from the Women's Interagency HIV Study in an HIV disease simulation model to estimate life expectancy in women who receive an efavirenz‐based initial antiretroviral regimen compared with those who delay efavirenz use and receive a boosted protease inhibitor‐based initial regimen. To estimate excess risk of teratogenic events with and without efavirenz exposure per 100 000 women, we incorporated literature‐based rates of pregnancy, live births, and teratogenic events into a decision analytic model. We assumed a teratogenicity risk of 2.90 events/100 live births in women exposed to efavirenz during pregnancy and 2.68/100 live births in unexposed women.

Results

Survival for HIV‐infected women who received an efavirenz‐based initial antiretroviral therapy (ART) regimen was 0.89 years greater than for women receiving non‐efavirenz‐based initial therapy (28.91 vs. 28.02 years). The rate of teratogenic events was 77.26/100 000 exposed women, compared with 72.46/100 000 unexposed women. Survival estimates were sensitive to variations in treatment efficacy and AIDS‐related mortality. Estimates of excess teratogenic events were most sensitive to pregnancy rates and number of teratogenic events/100 live births in efavirenz‐exposed women.

Conclusions

Use of non‐efavirenz‐based initial ART in HIV‐infected women of childbearing age may reduce life expectancy gains from antiretroviral treatment, but may also prevent teratogenic events. Decision‐making regarding efavirenz use presents a trade‐off between these two risks; this study can inform discussions between patients and health care providers.     

Lymphocytes as internal standard in oxidative burst analysis by cytometry: a new data analysis approach

We propose a new data analysis approach for reactive oxygen species detection using Dihydrorhodamine 123 in blood monocytes and neutrophils. This approach, based on data transformation using lymphocytes as internal standard, allows to appreciate free radical production by monocytes also without Phorbol 12-myristate 13-acetate (PMA) activation. In addition, this method is sensitive to differences in healthy subjects due to sub-pathological conditions, such as hypercholesterolemia.     

Helicobacter pylori regulates iNOS promoter by histone modifications in human gastric epithelial cells

Inducible nitric oxide synthase (iNOS) expression is altered in gastrointestinal diseases. Helicobacter pylori (Hp) infection may have a critical role in iNOS disregulation. We undertook this study to investigate possible chromatin changes occurring early during iNOS gene activation as a direct consequence of Hp-gastric cells interaction. We show that Hp infection is followed by different expression and chromatin modifications in gastric cells including (1) activation of iNOS gene expression, (2) chromatin changes at iNOS promoter including decreased H3K9 methylation and increased H3 acetylation and H3K4 methylation levels, (3) selective release of methyl-CpG-binding protein 2 from the iNOS promoter. Moreover, we show that Hp-induced activation of iNOS is delayed, but not eliminated, by the treatment with LSD1 inhibitors. Our data suggest a role for specific chromatin-based mechanisms in the control of human iNOS gene expression upon Hp exposure.