A non-coding cancer mutation disrupting an HNF4α binding motif affects an enhancer regulating genes associated to the progression of liver cancer.

BACKGROUND: Somatic mutations in coding regions of the genome may result in non-functional proteins that can lead to cancer or other diseases, however cancer mutations in the non-coding regions have rarely been studied and the interpretation of their effects is difficult. Non-coding mutations might act by breaking or creating transcription factor binding motifs in promoters, enhancers or silencers resulting in altered expression of target gene(s). A high number of mutations have been reported in coding and non-coding regions in cells of liver cancer. Hepatocyte nuclear factor 4α is a transcription factor that regulates the expression of several genes in liver cells, while the motifs it binds are frequently mutated in promoters and enhancers in liver cancer. AIM: The aim of the study is to evaluate the genetic effects of a non-coding somatic mutation frequently observed in liver cancer. MATERIALS AND METHODS: We evaluated experimentally the effects of a somatic mutation frequently reported in liver cancer as a motif-breaker for the binding of hepatocyte nuclear factor 4α. The effects of the mutation on protein binding and enhancer activity were studied in HepG2 cells via electrophoresis mobility shift assay and dual luciferase reporter assays. We also studied genome-wide promoter-enhancer interactions performing targeted chromosome conformation capture in liver tissue to identify putative target genes whose expression could be altered by the mutation. RESULTS: We found that the mutation leads to reduced protein binding and a decrease in enhancer activity. The enhancer harboring the mutation interacts with the promoters of ANAPC13, MAP6D1 and MUC13, which have been implicated in liver cancer. CONCLUSIONS: The study highlights the importance of non-coding somatic mutations, vastly understudied, but likely to contribute to cancer development and progression.

CD4- and CD8-expressing cells found in the bovine and porcine anterior chamber of the eye.

The aim of the present study was to investigate whether the anterior chamber constitutes part of the normal migratory pathway of CD4+ and CD8+ lymphocytes in cattle and swine. The cells obtained from aqueous humor of cows and pigs were stained for CD4 and CD8 receptors, and subsequently analyzed with flow cytometry. The mean percentage of CD4+CD8-, CD4-CD8+ and CD4+CD8+ cells within the total lymphocyte population of the bovine anterior chamber was, respectively, 17.88, 12.64 and 27.26%. In turn, the mean values of these parameters in pigs were 1.77, 38.48 and 17.45, respectively. Among bovine and porcine CD4+CD8+ cells prevalent were those displaying CD4lowCD8low and CD4lowCD8high phenotypes, respectively. The results suggest that the anterior chamber in cattle and swine is an element in the normal migratory pathway of CD4+, CD8+ and CD4+CD8+ cells. Furthermore, the contribution of these subsets in the anterior chamber lymphocyte population can differ considerably between animal species.

"Extracorporeal Membrane Oxygenation for Greater Poland" Program: How to Save Lives and Develop Organ Donation?

The "ECMO for Greater Poland" program takes full advantage of the extracorporeal membrane oxygenation (ECMO) perfusion therapy opportunities to promote the health of the 3.5 million inhabitants in the region. The main implementation areas are treatment of patients with hypothermia; severe reversible respiratory failure (RRF); critical states resulting in heart failure, that is, cardiac arrest, cardiogenic shock, or acute intoxication; and promotion of the donor after circulatory death (DCD) strategy in selected organ donor cases, after unsuccessful life-saving treatment, to achieve organ recovery. This organizational model is complex and expensive, so we used advanced high-fidelity medical simulation tests to prepare for real-life experience. Over the course of 4 months we performed scenarios including "ECMO for DCD," "ECMO for extended cardiopulmonary resuscitation," "ECMO for RRF," and "ECMO in hypothermia." Soon after these simulations, Maastricht category II DCD procedures were performed involving real patients and resulting in 2 successful double kidney transplantations for the first time in Poland. One month later we treated 2 hypothermia patients (7 adult patients with heart failure and 5 patients with reversible respiratory failure) with ECMO for the first time in the region. Fortunately, we have discovered an important new role of medical simulation. It can be used not only for skills testing but also as a tool to create non-existing procedures and unavailable algorithms. The result of these program activities will promote the care and treatment of patients in critical condition with ECMO therapy as well as increase the potential organ pool from DCDs in the Greater Poland region of Poland.

Effect of deoxynivalenol on the levels of toll-like receptors 2 and 9 and their mRNA expression in enterocytes in the porcine large intestine: a preliminary study.

Deoxynivalenol (DON), one of the most prevalent mycotoxins in the world, and is capable of inducing immune disorders in humans and animals. The aim of this study was to determine the effect of feed contaminated with DON on the number of TLR2- and TLR9-positive cells and their mRNA expression in the porcine large intestine. The experiment was conducted on two equal groups of pigs (n=4). The experimental group (E) was administered feed contaminated with DON (1008 µg/kg of feed) for 6 weeks, and the control group (C) was administered non-contaminated feed over the same period of time. A decrease in the expression of TLR2 mRNA was noted in the cecum. The percentage of TLR9-positive enterocytes increased in the ascending colon and decreased in the cecum. The results of this study indicate that DON can modify the local immune response by changing the expression of TLRs.

An analysis of factors affecting the mercury content in the human femoral bone.

The study was carried out to determine the content of mercury in bone tissue of the proximal femur (head and neck bone) of 95 patients undergoing total hip replacement due to osteoarthritis, using CF-AFS analytical technique. Furthermore, the investigations were aimed at assessing the impact of selected factors, such as age, gender, tobacco smoking, alcohol consumption, exposure to chemical substance at work, type of degenerative changes, clinical evaluation and radiological parameters, type of medications, on the concentration of mercury in the head and neck of the femur, resected in situ. Mercury was obtained in all samples of the head and neck of the femur (n = 190) in patients aged 25-91 years. The mean content of mercury for the whole group of patients was as follows: 37.1 ± 35.0 ng/g for the femoral neck and 24.2 ± 19.5 ng/g for the femoral head. The highest Hg contents were found in femoral neck samples, both in women and men, and they amounted to 169.6 and 176.5 ng/g, respectively. The research showed that the mercury content of bones can be associated with body mass index, differences in body anatomy, and gender. The uses of statistical analysis gave the possibility to define the influence of factors on mercury content in human femoral bones.

The role of sentinel node mapping with indocyanine green and endoscopic near-infrared fluorescence imaging in endometrial and cervical cancer.

INTRODUCTION: Endometrial and cervical carcinoma are common neoplasms in gynecological oncology. The prognosis and treatment depend on the stage of the cancer according to the FIGO staging system. Stage IAl may be treated by hysterectomy or even local surgical procedures. For Stage IA2, radical hysterectomy and lymphadenectomy must be performed. Lymph node metastasis is an important prognostic factor in both cancers, however lymphadenectomy is associated with long-term complications. Thanks to the sentinel lymph node biopsy (SLNB), we can more accurately discover the staging of the primary tumor, and in case of sentinel lymph node (SLN) negative patients, can resign regional lymphadenectomy. Some researchers claim that new techniques such as indocyanine green (ICG) and endoscopic near-infrared fluorescence imaging for sentinel node mapping can be used instead of the traditional techniques. AIM: To establish the role of sentinel node mapping technique in endometrial and cervical cancer. MATERIAL AND METHODS: A retrospective study of medical records of five patients with cervical cancer (first group) Stage I and nine patients (second group) who underwent laparoscopic radical hysterectomy and SLNB or group of lymph nodes. These procedures were performed at Gynecology Department of the District Hospital in Garwolin. RESULTS: All lymph nodes were clear of metastases. All patients after histopathological diagnosis were finally referred to the Cancer Centre and Institute of Oncology due to consultation or for further treatment. CONCLUSION: Based on the present first results and literature review, intracervical ICG injection with fluorescence imaging seems to be the best SLN mapping technique, because of its simplicity, safety, and overall lower cost. More data is required to determine if the nodes identified with this technique are able to predict metastatic disease.

Magnetically tuned, robust and efficient filtering system for spatially multimode quantum memory in warm atomic vapors.

Warm atomic vapor quantum memories are simple and robust, yet suffer from a number of parasitic processes which produce excess noise. For operating in a single-photon regime precise filtering of the output light is essential. Here, we report a combination of magnetically tuned absorption and Faraday filters, both light-direction insensitive, which stop the driving lasers and attenuate spurious fluorescence and four-wave mixing while transmitting narrowband Stokes and anti-Stokes photons generated in write-in and readout processes. We characterize both filters with respect to adjustable working parameters. We demonstrate a significant increase in the signal-to-noise ratio upon applying the filters seen qualitatively in measurements of correlation between the Raman scattered photons.

Tumour-processed osteopontin and lactadherin drive the protumorigenic reprogramming of microglia and glioma progression.

Tumour tissue is infiltrated by myeloid cells that are reprogrammed into alternatively activated/regenerative (M2) macrophages. The contribution of major signalling pathways and their modulators/targets involved in the macrophage reprogramming is poorly known. Glioblastoma (malignant brain tumour) attracts and reprograms brain-resident microglia and peripheral macrophages into cells that increase invasion, angiogenesis and suppress antitumour immunity. Using a 'function-first' approach and glioma secretome proteomics we identified osteopontin and lactadherin as proteins that cooperatively activate amoeboid transformation, phagocytosis and motility of primary microglia cultures via integrins and FAK-Akt (focal adhesion kinase-Akt) signalling. A synthetic peptide interfering with integrin ligands blocks glioma-microglia communication, functional activation and M2 gene expression. We found that osteopontin/secreted phosphoprotein 1 (Spp1) produced by non-transformed cells acts as a proinflammatory factor inducing inflammatory signalling and M1 genes, and counteracts the action of lactadherin. Using constructs encoding functional mutants of osteopontin, we demonstrated sequential processing of Spp1 by thrombin and matrix metalloproteinase-3 and/or -7 (MMP-3 and/or -7) in glioma cells, which generates a microglia-activating form devoid of the inflammatory activity, while retaining the M2 reprogramming potential. A similar form of osteopontin is secreted by human glioma cells but not normal human astrocytes. Knockdown of osteopontin or lactadherin in glioma cells reduces intracranial glioma growth, blocks amoeboid transformation of myeloid cells and affects M2 reprogramming of microglia/macrophages. Our findings demonstrate how glioma cells misuse macrophage-activating signals and redesign primarily proinflammatory signals towards their advantage to induce M2 reprogramming of tumour-infiltrating brain macrophages.

Nitrogen-boron coordination versus OH∙∙∙N hydrogen bonding in pyridoxaboroles - aza analogues of benzoxaboroles.

Pyridoxaboroles - fused heterocyclic systems composed of pyridine and five-membered oxaborole rings - have been obtained for the first time from simple halopyridines. Thus, 6-butyl-2-(3'-bromo-4'-pyridyl)-(N-B)-1,3,6,2-dioxazaborocan obtained from 3-bromopyridine was converted into a lithio derivative by Br/Li exchange using nBuLi/THF at -85 °C. This intermediate was trapped with benzaldehydes to give the corresponding pyridoxaboroles after hydrolysis. The use of chlorodiphenylsilane as an electrophile gave rise to a related pyridosiloxaborole. The fluorinated pyridoxaborole was obtained by deprotonation of α-(2-methoxyphenyl)-2-fluoro-4-iodopyridylmethanol with NaH and consecutive iodine-lithium exchange/boronation followed by hydrolysis. Single-crystal X-ray analysis of pyridino[4,3-c]-1,3-dihydro-1-hydroxy-3-mesityl[2,1]oxaborole revealed the formation of a unique 1D coordination polymer based on N-B dative bonds between monomeric molecules. In contrast, the crystal structure of 2-fluoropyridino[4,3-c]-1,3-dihydro-1-hydroxy-3-(2'-methoxyphenyl)[2,1]oxaborole features an infinite H-bonded chain as the main structural motif. The presented considerations are quantified in terms of various computational methods (single molecule and dimer energy calculations, electron density topology, NBO analyses) providing a comprehensive picture of the structural properties of pyridoxaboroles.

'True' null allele detection in microsatellite loci: a comparison of methods, assessment of difficulties and survey of possible improvements.

Null alleles are alleles that for various reasons fail to amplify in a PCR assay. The presence of null alleles in microsatellite data is known to bias the genetic parameter estimates. Thus, efficient detection of null alleles is crucial, but the methods available for indirect null allele detection return inconsistent results. Here, our aim was to compare different methods for null allele detection, to explain their respective performance and to provide improvements. We applied several approaches to identify the 'true' null alleles based on the predictions made by five different methods, used either individually or in combination. First, we introduced simulated 'true' null alleles into 240 population data sets and applied the methods to measure their success in detecting the simulated null alleles. The single best-performing method was ML-NullFreq_frequency. Furthermore, we applied different noise reduction approaches to improve the results. For instance, by combining the results of several methods, we obtained more reliable results than using a single one. Rule-based classification was applied to identify population properties linked to the false discovery rate. Rules obtained from the classifier described which population genetic estimates and loci characteristics were linked to the success of each method. We have shown that by simulating 'true' null alleles into a population data set, we may define a null allele frequency threshold, related to a desired true or false discovery rate. Moreover, using such simulated data sets, the expected null allele homozygote frequency may be estimated independently of the equilibrium state of the population.

Recipient uridine 5'-diphospho-glucuronosyltransferase UGT1A9 c.98T>C variant determines transplanted kidney filtration rate.

Mycophenolic acid preparations are commonly used in prophylaxis of kidney allograft acute rejection. The medication is metabolized by uridine diphosphate glucuronosyltransferases, mainly UGT1A9 present in liver, kidney, and intestine. The effect of UGT1A9 allelic variants on drug metabolism in healthy volunteers and transplant recipients has been previously evaluated; these studies included the UGT1A9 c.98T>C polymorphism (rs72551330, p.Met33Thr) causing methionine-to-threonine substitution in the polypeptide chain. The study objective was to evaluate the relationship between UGT1A9 c.98T>C polymorphism and kidney graft function and survival and the risk of acute allograft rejection. Kidney recipients who underwent transplantation between 2000 and 2007 at the Medical University of Warsaw were included. Clinical data originated from standard medical records, UGT1A9 c.98T>C was genotyped using a polymerase chain reaction-restriction fragment length polymorphism method. A group of 243 kidney transplant recipients was enrolled in the study. The frequency of the c.98C allele was 2.4% (12 of 486). Most of the carriers of the allelic variant (10 of 12) received cyclosporine A at transplantation. In the c.98C allele carriers, worse function of the renal allograft, manifested by a significantly lower glomerular filtration rate, was found in the first posttransplantation month and persisted at a lower level for 8 years after the procedure (for comparison of the UGT1A9 c.98 TT vs the UGT1A9 c.98 TC groups, P = .03). No association was found between the presence of the UGT1A9 c.98C allele and the risk of delayed renal graft function, acute rejection, dysfunction of the renal graft, or dialysis treatment reintroduction. Significantly lower estimated glomerular filtration rate of the renal allograft in UGT1A9 c.98C carriers did not translate into decreased allograft survival.

Oscillations of the orbital magnetic moment due to d-band quantum well states.

The effect of electron confinement on the magnetocrystalline anisotropy of ultrathin bcc Fe films is explored by combining photoemission spectroscopy, x-ray magnetic circular dichroism, and magneto-optical Kerr effect measurements. Pronounced thickness-dependent variations in the magnetocrystalline anisotropy are ascribed to periodic changes in the density of states at the Fermi level, induced by quantization of d(xz), d(yz) out-of-plane orbitals. Our results reveal a direct correlation between quantum well states, the orbital magnetic moment, and the magnetocrystalline anisotropy.

The outcome of care in people with type 1 diabetes after switching to insulin glargine-based regimens in a real-life setting: a long-term observational study.

BACKGROUND: Different insulin preparations are used as basal insulins in type 1 diabetes. The aim of this study was to assess long-term efficacy and safety of insulin glargine after switching from other basal insulins in type 1 diabetes in a real-life setting. METHODS: In the clinic's database, 87 subjects treated with glargine for > 1 year were identified. In all patients, HbA1c level, insulin doses, episodes of severe hypoglycaemia, diabetic complications, comorbidities, body mass index (BMI), blood pressure and concomitant medications' use were monitored throughout the entire follow-up period. RESULTS: During observation, lasting mean 61.9 ± 27.6 months HbA1c level decreased from 8.86 ± 1.60% (73.3 mmol/mol) to 8.25 ± 1.40% (66.7 mmol/mol), p < 0.001. This improvement was maintained up to 8 years. Frequency of severe hypoglycaemia was 6.24/100 patient-years. Total insulin requirement did not changed significantly. BMI increased from 23.57 ± 2.90 to 24.52 ± 3.46 kg/m(2) (p < 0.001). Significant weight gain (> 5%) occurred in 30 subjects, while 10 patients lost weight. Mean systolic blood pressure (SBP) increased from 136.3 ± 13.4 to 140.7 ± 15.1 mmHg (p = 0.008), while diastolic blood pressure remained unchanged. Development or progression of diabetic complications was revealed in 11 subjects. CONCLUSIONS: Following switch from other basal insulins to insulin glargine in type 1 diabetic patients, glycaemic control significantly improved, with unchanged total insulin requirement and with low risk of severe hypoglycaemia. Weight gain and elevation of SBP observed in this study require special attention and educational efforts. In summary, insulin glargine can be recommended as an effective and safe basal insulin in type 1 diabetes in a real-life setting.

Reliability assessment of null allele detection: inconsistencies between and within different methods.

Microsatellite loci are widely used in population genetic studies, but the presence of null alleles may lead to biased results. Here, we assessed five methods that indirectly detect null alleles and found large inconsistencies among them. Our analysis was based on 20 microsatellite loci genotyped in a natural population of Microtus oeconomus sampled during 8 years, together with 1200 simulated populations without null alleles, but experiencing bottlenecks of varying duration and intensity, and 120 simulated populations with known null alleles. In the natural population, 29% of positive results were consistent between the methods in pairwise comparisons, and in the simulated data set, this proportion was 14%. The positive results were also inconsistent between different years in the natural population. In the null-allele-free simulated data set, the number of false positives increased with increased bottleneck intensity and duration. We also found a low concordance in null allele detection between the original simulated populations and their 20% random subsets. In the populations simulated to include null alleles, between 22% and 42% of true null alleles remained undetected, which highlighted that detection errors are not restricted to false positives. None of the evaluated methods clearly outperformed the others when both false-positive and false-negative rates were considered. Accepting only the positive results consistent between at least two methods should considerably reduce the false-positive rate, but this approach may increase the false-negative rate. Our study demonstrates the need for novel null allele detection methods that could be reliably applied to natural populations.

Low risk of severe hypoglycaemia in patients with type 2 diabetes mellitus starting insulin therapy with premixed insulin analogues BID in outpatient settings.

AIMS: The choice of insulin at initiation in type 2 diabetes remains controversial. The aim of this study was to assess the occurrence of self-reported severe hypoglycaemia associated with premixed insulin analogues in routine clinical care. METHODS: A 12-month, prospective, observational, multicentre study in patients starting a commonly prescribed premixed insulin analogue (either insulin lispro 25/75 or biphasic insulin aspart 30/70, twice daily) after suboptimal glycaemic control on oral antidiabetic agents. Treatment decisions were made solely in the course of usual practice. RESULTS: Study follow-up was completed by 991 (85.5%) of the 1150 patients enrolled. At baseline, mean (SD) age was 57.9 (10.1) years; mean diabetes duration was 9.2 (5.9) years; mean haemoglobin A(1c) (HbA(1c)) was 9.9 (1.8) % and the rate of severe hypoglycaemia was 0.03 episode/patient-year. At 12 months, the rate of severe hypoglycaemia was 0.04 episode/patient-year (95% CI 0.023, 0.055 episode/patient-year) and mean insulin dose was 41.5 (19.4) units. Changes from baseline to 12 months for mean fasting plasma glucose and HbA(1c) were -5.1 mmol/l and -2.5%, respectively. CONCLUSIONS: After initiation of premixed insulin analogues in patients with type 2 diabetes in real-world settings, the incidence of severe hypoglycaemia was lower than expected from previously reported studies.

Osteoblastoma of the nasal septum.

OBJECTIVES: To present a case of, and to review the literature concerning, osteoblastoma of the nasal cavity, and to demonstrate the importance of considering this rare entity when assessing patients presenting with a nasal septum lesion. CASE REPORT: Benign osteoblastoma is a rare tumour, constituting 1 per cent of all bone tumours. Most cases occur in the long bones. Osteoblastoma involving the nasal cavity is rare, with only 10 reported cases in the English-language literature. Most nasal cavity cases originate from the ethmoid sinus and spread to involve the nasal cavity. There are only four reported cases of osteoblastoma originating from the bones of the nasal cavity. We report a case of osteoblastoma originating from the bony nasal septum in a 45-year-old man with a history of recurrent, right-sided epistaxis and nasal obstruction. CONCLUSION: This is the second report in the English-language literature of osteoblastoma originating from the bony nasal septum.

Dose-dependent in vivo effect of Rhodiola and Echinacea on the mitogen-induced lymphocyte proliferation in mice.

Echinacea purpurea (EP) and Echinacea angustifolia (EA) are ones of the most important world's herbs with immunotropic activity. They were traditional medicinal plants used by North American Indians for the treatment of various illnesses. Now they are cultivated in many countries and are used mainly to treat respiratory tract infections. Rhodiola rosea (RR) and Rhodiola quadrifida (RQ) are medicinal plants originated from Asia and used traditionally as adaptogens, antidepressants, and anti-inflammatory remedies. We previously reported, that extracts of underground parts of RR and RQ exhibited immunotropic activity. We have demonstrated in pigs that in vitro RR or RQ supplementation of blood lymphocyte cultures stimulated T cell proliferative response to Con A in lower, and inhibited it in higher Rhodiola extract concentrations. The aim of this work was to evaluate the in vivo effect of these herbal remedies on the in vitro proliferative response of mouse splenic lymphocytes to another T-cell mitogen- Phaseolus vulgaris haemagglutinin (PHA). We have found significant stimulation of proliferative response, in comparison to the controls, in mice fed lower doses of tested remedies, and inhibition, no effect or lower stimulation, in mice fed higher doses of these drugs.

Analysis of expression and structure of the TSG101 gene in cervical cancer cells.

Human papillomavirus (HPV)-mediated transformation of human epithelial cells has been recognized as a multi-step process in which additional unknown factors and (epi)genetic events are required. The tumor susceptibility gene 101 (TSG101) was discovered in mouse NIH3T3 fibroblast cells as a gene whose functional knockout leads to transformation. TSG101 protein is involved in a variety of important biological functions, such as ubiquitination, transcriptional regulation, endosomal trafficking, virus budding, proliferation and cell survival. It is suggested that TSG101 is an important factor for maintaining cellular homeostasis and that perturbation of TSG101 functions leads to cell transformation. Interestingly, a recent report showed up- or down-regulation of TSG101 in several human malignancies. At present, the role of TSG101 in cervical tumorigenesis is unexplained. TSG101 expression in tumors, where carcinogenesis is connected with viral infection, and a mechanism of TSG101 expression regulation in cancer cells are also unknown. The aim of our study was to estimate the TSG101 mRNA and protein level in cervical cancer and non-tumor epithelial cells. We also analyzed the TSG101 coding and promoter sequence using the PCR-SSCP technique and methylation pattern of the TSG101 promoter. Our real-time PCR and Western blot analysis showed decreased TSG101 mRNA and protein level in cervical cancer tissue in comparison to normal (non-tumor) HPV(-) and HPV16(+) epithelial cells. Our results suggest that TSG101 down-regulation in cervical cancer cells is not regulated by genetic or epigenetic events. However, we detected novel single nucleotide polymorphisms in the promoter of this gene.

Reduced effect of propofol at human {alpha}1{beta}2(N289M){gamma}2 and {alpha}2{beta}3(N290M){gamma}2 mutant GABA(A) receptors.

BACKGROUND: Propofol is an i.v. anaesthetic commonly used during general anaesthesia and intensive care. It is known that the second transmembrane segment of the beta subunit in the GABA(A) receptor is an important target for the effects of propofol; however, this has not been investigated in human receptors. The aim of this study was to investigate the effect of propofol on human beta2 and beta3 GABA(A) subunits with point mutations corresponding to the N265M mutation in the rat beta2 and beta3 subunits. METHODS: Asparagine-to-methionine replacement at amino acid position 289 and 290 (N289M and N290M) in the beta2 and beta3 GABA(A) receptor subunits, respectively, was accomplished by site-directed mutagenesis. Thereafter, subunits for three human wild-type (alpha1beta2gamma2, alpha2beta2gamma2, and alpha2beta3gamma2) and two mutant GABA(A) receptor channels [alpha1beta2(N289M)gamma2 and alpha2beta3(N290M)gamma2] were introduced into Xenopus oocytes and studied with two-electrode voltage clamp. RESULTS: The mutant receptors left-shifted the GABA concentration-response curve. In comparison with the wild-type receptors, both the positive modulatory and the agonistic effects of propofol were strongly reduced in potency and amplitude at both mutated GABA(A) channels. CONCLUSIONS: We demonstrate that N289M or N290M mutation in human GABA(A) beta2 and beta3 subunits increases sensitivity to GABA, which is in contrast to the corresponding rat N265M mutation. Furthermore, the N289M and N289M mutations reduce both the potentiation of GABA-induced currents and the direct effect of propofol on channels incorporating either of the mutated subunits, which confirms earlier findings concerning the corresponding mutation in rat receptors and knock-in mice.