Predictors of cervical cancer screening service utilization among female health care workers in a tertiary hospital in Osogbo, Osun State.

Cervical cancer is the second commonest cancer in Nigerian women that can be reduced through effective screening. This study aimed to determine the utilization of cervical cancer screening services among female health care workers in a tertiary hospital. It is a descriptive cross-sectional study that utilized a pretested questionnaire to collect data from 170 respondents. Data were analyzed using descriptive statistics and logistic regression. 61.2% had good knowledge about cervical cancer, and 75.3% reported utilization of cervical cancer screening services. Factors that affect utilization of cervical cancer screening services are fear of negative results (82.4%), lack of time (78.2%), cost of screening (63.5%), and support from husband (61.2%). Logistic regression analysis showed that age (ß = 4.009, p = < 0.001), year of experience (ß = -4.350, p <0.001), fear of negative results (ß = -2.479, p = 0.014) and lack of support from husband (ß = 4.380, p < 0.001) significantly predict utilization of screening services. Conclusively, female health workers know about the prevention of cervical cancer and are willing to utilize screening. It is recommended that cervical cancer screening should be made mandatory for female health care workers, to reduce the incidence of cervical cancer cases.

Orai1 and Orai2 mediate store-operated calcium entry that regulates HL60 cell migration and FAK phosphorylation.

Store-operated Ca2+ entry (SOCE) is a major mechanism for the regulation of intracellular Ca2+ homeostasis and cellular function. Emerging evidence has revealed that altered expression and function of the molecular determinants of SOCE play a critical role in the development or maintenance of several cancer hallmarks, including enhanced proliferation and migration. Here we show that, in the acute myeloid leukemia cell line HL60, Orai2 is highly expressed at the transcript level, followed by the expression of Orai1. Using fluorescence Ca2+ imaging we found that Orai2 silencing significantly attenuated thapsigargin-induced SOCE, as well as knockdown of Orai1, while silencing the expression of both channels almost completely reduced SOCE, thus suggesting that SOCE in these cells is strongly dependent on Orai1 and Orai2. On the other hand, the expression of TRPC1, TRPC3 and TRPC6 is almost absent at the transcript and protein level. Bromodeoxyuridine cell proliferation assay revealed that Orai1 and Orai2 expression silencing significantly reduced HL60 cell proliferation. Furthermore, knockdown of Orai1 and Orai2 significantly attenuated the ability of HL60 to migrate in vitro as determined by transwell migration assay, probably due to the impairment of FAK tyrosine phosphorylation. These findings provide evidence for a role for Orai1 and Orai2, in SOCE and migration in the human HL60 promyeloblastic cell line. This article is part of a Special Issue entitled: ECS Meeting edited by Claus Heizmann, Joachim Krebs and Jacques Haiech.

Control of H_{2} Dissociative Ionization in the Nonlinear Regime Using Vacuum Ultraviolet Free-Electron Laser Pulses.

The role of the nuclear degrees of freedom in nonlinear two-photon single ionization of H_{2} molecules interacting with short and intense vacuum ultraviolet pulses is investigated, both experimentally and theoretically, by selecting single resonant vibronic intermediate neutral states. This high selectivity relies on the narrow bandwidth and tunability of the pulses generated at the FERMI free-electron laser. A sustained enhancement of dissociative ionization, which even exceeds nondissociative ionization, is observed and controlled as one selects progressively higher vibronic states. With the help of ab initio calculations for increasing pulse durations, the photoelectron and ion energy spectra obtained with velocity map imaging allow us to identify new photoionization pathways. With pulses of the order of 100 fs, the experiment probes a timescale that lies between that of ultrafast dynamical processes and that of steady state excitations.

Does Lung Donation by Heart Donors Have an Impact on Survival in Heart Transplant Recipients?

Lung procurement is increasing during multiorgan recovery and substantially alters the explant process. This study evaluated whether lung donation by a heart donor affects survival in heart transplant recipients. Retrospective analysis of United Network for Organ Sharing (UNOS) adult heart transplantation data from 1998 to 2012 was performed. Lung donors (LDs) were defined as those having at least one lung procured and transplanted. Non-LDs had neither lung transplanted. Heart transplant recipients who had previous transplants, who had heterotopic transplants, who were waitlisted for other organs or who were temporarily delisted were excluded from the analysis. Kaplan-Meier survival analysis and Cox proportional hazards regression were performed. Of 23 590 heart transplant recipients meeting criteria during the study period, 8638 (36.6%) transplants were from LDs. Donors in the LD group had less history of cigarette use (15.5% vs. 29.5%, p < 0.001). On univariate analysis, LDs were associated with improved patient survival (p < 0.001). On multivariate analysis, LDs were not significantly associated with patient survival (adjusted hazard ratio 0.98, 95% confidence interval 0.94-1.03). Analysis of the UNOS registry suggested that donor pulmonary status and lung procurement had no detrimental effect on survival in heart transplant recipients, supporting the present practice of using donor lungs whenever possible.

A comparison of the mandibular index on panoramic and cross-sectional images from CBCT exams from osteoporosis risk group.

UNLABELLED: Osteoporosis is a degenerative disease that primarily affects postmenopausal women. Based on panoramic radiographs, several assessment methods have been proposed for the diagnosis and evaluation of bone changes and as a predictor of osteoporosis for example the mandibular index. INTRODUCTION: The purpose of this study is to compare the assessment of mandibular indices on panoramic and cross-sectional images. METHODS: Forty-four cone beam computed tomography (CBCT) images from postmenopausal female subjects aged more than 45 years without systemic changes were selected for this study. From those images, cross-sectional and panoramic reconstruction images were assembled into a template for evaluation. The evaluation was conducted by observing the panoramic images and parasagittal sections. The appearance of the inferior cortex of the mandible was classified according to the mandibular index: C1, the endosteal margin of the cortex was even and sharp; C2, the endosteal margin presented semilunar defects or appeared to form endosteal cortical residues; or C3, the cortical layer formed heavy endosteal cortical residues and was clearly porous. RESULTS: Based on Wilcoxon statistical test (p > 0.01), the data showed no statistically significant difference between the exams. CONCLUSION: The mandibular index assigned in tomographic images is comparable to that obtained in panoramic images, indicating a valid use of the index in CBCT images, which can lead to the identification of patients with bone mass loss and a premature referral to further exams and treatment.

Dosimetry of ultra-high dose rate electron beams using thermoluminescence and optically stimulated luminescence detectors.

Objective.The aim of this work is to investigate the dose rate dependence of thermoluminescence and optically stimulated luminescence detectors (TLDs and OSLDs) in a wide uniform ultra-high dose rate electron beam and demonstrate the potential use of TLDs and OSLDs to correct the ion recombination in air-filled ionization chambers. This study avoids previously reported complications related to the field size and homogeneity.Approach.Two types of OSLDs (BeO and Al2O3:C) and three types of TLDs (LiF:Mg,Ti, LiF:Mg,Cu,P, CaF2:Tm) were irradiated simultaneously in a uniform 16 MeV electron beam generated by a clinically decommissioned C-Arm LINAC, modified to deliver doses per pulse between 8.3 × 10-4Gy and 1.255 Gy, corresponding to instantaneous dose rates between 2 × 102Gy s-1and 3 × 105Gy s-1. A prototype ultra-thin parallel plate ionization chamber was employed as reference detector.Main results.Reproducible results were achieved both at conventional (standard deviation of the data <2%) and at the highest dose per pulse (standard deviation of the data <4%). No trend in the dose rate response of the TLDs and OSLDs was observed in the investigated dose per pulse range. The Al2O3:C OSLD was found to be the most precise detector, with a standard deviation of the data <2% at all investigated dose rates and dose levels.Significance.The dose rate independence of the investigated TLDs and OSLDs make them good candidates for dosimetry at ultra-high dose rates, at least up to 3 × 105Gy s-1. A dose rate independent method to measure the dose per pulse is proposed, which can be applied to characterize ultra-high dose rate electron beams and correct for ion recombination in ionization chambers.

Results of 2023 survey on the use of synthetic computed tomography for magnetic resonance Imaging-only radiotherapy: Current status and future steps.

Background and purpose: The emergence of synthetic CT (sCT) in MR-guided radiotherapy (MRgRT) represents a significant advancement, supporting MR-only workflows and online treatment adaptation. However, the lack of consensus guidelines has led to varied practices. This study reports results from a 2023 ESTRO survey aimed at defining current practices in sCT development and use. Materials and methods: An survey was distributed to ESTRO members, including 98 questions across four sections on sCT algorithm generation and usage. By June 2023, 100 centers participated. The survey revealed diverse clinical experiences and roles, with primary sCT use in the pelvis (60%), brain (15%), abdomen (11%), thorax (8%), and head-and-neck (6%). sCT was mostly used for conventional fractionation treatments (68%), photon SBRT (40%), and palliative cases (28%), with limited use in proton therapy (4%). Results: Conditional GANs and GANs were the most used neural network architectures, operating mainly on 1.5 T and 3 T MRI images. Less than half used paired images for training, and only 20% performed image selection. Key MR image quality parameters included magnetic field homogeneity and spatial integrity. Half of the respondents lacked a dedicated sCT-QA program, and many did not apply sanitychecks before calculation. Selection strategies included age, weight, and metal artifacts. A strong consensus (95%) emerged for vendor neutral guidelines. Conclusion: The survey highlights the need for expert-based, vendor-neutral guidelines to standardize sCT tools, metrics, and clinical protocols, ensuring effective sCT use in MR-guided radiotherapy.

High genetic diversity of Newcastle disease virus in poultry in West and Central Africa: cocirculation of genotype XIV and newly defined genotypes XVII and XVIII.

Despite rampant Newcastle disease virus (NDV) outbreaks in Africa for decades, the information about the genetic characteristics of the virulent strains circulating in West and Central Africa is still scarce. In this study, 96 complete NDV fusion gene sequences were obtained from poultry sampled in Cameroon, Central African Republic, Côte d'Ivoire, and Nigeria between 2006 and 2011. Based on rational criteria recently proposed for the classification of NDV strains into classes, genotypes, and subgenotypes, we revisited the classification of virulent strains, in particular those from West and Central Africa, leading to their grouping into genotype XIV and newly defined genotypes XVII and XVIII, each with two subgenotypes. Phylogenetic analyses revealed that several (sub)genotypes are found in almost every country. In Cameroon, most strains were related to vaccine strains, but a single genotype XVII strain was also found. Only three highly similar genotype XVII strains were detected in Central African Republic. Subgenotypes XVIIa, XVIIIa, and XVIIIb cocirculated in Côte d'Ivoire, while subgenotypes XIVa, XIVb, XVIIa, XVIIb, and XVIIIb were found in Nigeria. While these genotypes are so far geographically restricted, local and international trade of domestic and exotic birds may lead to their spread beyond West and Central Africa. A high genetic diversity, mutations in important neutralizing epitopes paired with suboptimal vaccination, various levels of clinical responses of poultry and wild birds to virulent strains, strains with new cleavage sites, and other genetic modifications found in these genotypes tend to undermine and complicate NDV management in Africa.

Genetic diversity of newcastle disease virus in wild birds and pigeons in West Africa.

In West and Central Africa, virulent Newcastle disease virus (NDV) strains of the recently identified genotypes XIV, XVII, and XVIII are enzootic in poultry, representing a considerable threat to the sector. The increasing number of reports of virulent strains in wild birds at least in other parts of the world raised the question of a potential role of wild birds in the spread of virulent NDV in sub-Saharan Africa as well. We investigated 1,723 asymptomatic birds sampled at live-bird markets and sites important for wild-bird conservation in Nigeria and 19 sick or dead wild birds in Côte d'Ivoire for NDV class I and II. Typical avirulent wild-type genotype I strains were found in wild waterfowl in wetlands in northeastern Nigeria. They were unrelated to vaccine strains, and the involvement of inter- or intracontinental migratory birds in their circulation in the region is suggested. Phylogenetic analyses also revealed that genotype VI strains found in pigeons, including some putative new subgenotype VIh and VIi strains, were introduced on multiple separate occasions in Nigeria. A single virulent genotype XVIII strain was found in a dead wild bird in Côte d'Ivoire, probably as a result of spillover from sick poultry. In conclusion, screening of wild birds and pigeons for NDV revealed the presence a variety of virulent and avirulent strains in West Africa but did not provide strong evidence that wild birds play an important role in the spread of virulent strains in the region.

Adaptive fractionation at the MR-linac.

Objective. Fractionated radiotherapy typically delivers the same dose in each fraction. Adaptive fractionation (AF) is an approach to exploit inter-fraction motion by increasing the dose on days when the distance of tumor and dose-limiting organs at risk (OAR) is large and decreasing the dose on unfavorable days. We develop an AF algorithm and evaluate the concept for patients with abdominal tumors previously treated at the MR-linac in 5 fractions.Approach. Given daily adapted treatment plans, inter-fractional changes are quantified by sparing factorsδtdefined as the OAR-to-tumor dose ratio. The key problem of AF is to decide on the dose to deliver in fractiont, givenδtand the dose delivered in previous fractions, but not knowing futureδts. Optimal doses that maximize the expected biologically effective dose in the tumor (BED10) while staying below a maximum OAR BED3constraint are computed using dynamic programming, assuming a normal distribution overδwith mean and variance estimated from previously observed patient-specificδts. The algorithm is evaluated for 16 MR-linac patients in whom tumor dose was compromised due to proximity of bowel, stomach, or duodenum.Main Results. In 14 out of the 16 patients, AF increased the tumor BED10compared to the reference treatment that delivers the same OAR dose in each fraction. However, in 11 of these 14 patients, the increase in BED10was below 1 Gy. Two patients with large sparing factor variation had a benefit of more than 10 Gy BED10increase. For one patient, AF led to a 5 Gy BED10decrease due to an unfavorable order of sparing factors.Significance. On average, AF provided only a small increase in tumor BED. However, AF may yield substantial benefits for individual patients with large variations in the geometry.

Immunodiagnosis of neurocysticercosis: ways to focus on the challenge.

Neurocysticercosis (NCC) is a disease of the central nervous system that is considered a public health problem in endemic areas. The definitive diagnosis of this disease is made using a combination of tools that include imaging of the brain and immunodiagnostic tests, but the facilities for performing them are usually not available in endemic areas. The immunodiagnosis of NCC is a useful tool that can provide important information on whether a patient is infected or not, but it presents many drawbacks as not all infected patients can be detected. These tests rely on purified or semipurified antigens that are sometimes difficult to prepare. Recent efforts have focused on the production of recombinant or synthetic antigens for the immunodiagnosis of NCC and interesting studies propose the use of new elements as nanobodies for diagnostic purposes. However, an immunodiagnostic test that can be considered as "gold standard" has not been developed so far. The complex nature of cysticercotic disease and the simplicity of common immunological assumptions involved explain the low scores and reproducibility of immunotests in the diagnosis of NCC. Here, the most important efforts for developing an immunodiagnostic test of NCC are listed and discussed. A more punctilious strategy based on the design of panels of confirmed positive and negative samples, the use of blind tests, and a worldwide effort is proposed in order to develop an immunodiagnostic test that can provide comparable results. The identification of a set of specific and representative antigens of T. solium and a thorough compilation of the many forms of antibody response of humans to the many forms of T. solium disease are also stressed as necessary.

Avoiding the trap: Mechanisms developed by pathogens to escape neutrophil extracellular traps.

Neutrophils are the first cells of the innate immune system that respond to infection by arriving at sites when pathogens have exceeded physical barriers. Among their response mechanisms against pathogens is the release of neutrophil extracellular traps (NETs), which are composed of deoxyribonucleic acid and antimicrobial proteins such as neutrophil elastase, myeloperoxidase, antimicrobial peptides, and other proteins in neutrophil granules. The formation of extracellular traps is considered an effective strategy to capture and, in some cases, neutralize pathogenic bacteria, fungi, parasites, or viruses. However, it is also known that pathogens can respond to NETs by expressing some virulence factors, thus evading the antimicrobial effect of these structures. These include the secretion of proteins to degrade the deoxyribonucleic acid scaffold, the formation of biofilms that impede the effect of NETs, or the modification of its membrane structure to avoid interaction with NETs. In this review, we discuss these mechanisms and summarize the different pathogens that employ one or more mechanisms to evade the NET-mediated neutrophil response.

Immunoendocrine host-parasite interactions during helminth infections: from the basic knowledge to its possible therapeutic applications.

Most of the current research on parasitic infections that affect humans and domestic animals has been focused on vaccines, diagnostic methods, epidemiology, new drug design, and recently, with the advancement of genomics and proteomics, on the evolutionary origins of parasites. However, the basic biology of many parasites of medical and veterinary importance has not been intensively studied. Some efforts have been made to obtain information on the parasite-host relationship; however, knowledge of the intricate neuroimmunoendocrine interactions of the host-parasite network, the consequences of this interaction on the host and parasite physiology, and its possible applications needs further investigation. We review here the literature, our own studies on the host-parasite neuroimmunoendocrine network, and how this basic knowledge can be used to design new treatments, by way of using hormones, antihormones, and hormone analogues as a possible novel therapy during parasitic diseases, with special emphasis on helminth parasites. Besides the biological interest, these investigations may contribute to the future identification of alternative treatments for several parasitic diseases. This complicated neuroimmunoendocrine network management during parasitic infections, and its physiological and behavioural consequences upon the host, may be operative in other mammalian infections. Such complexity may also help to explain the often conflicting results, observed between infections with respect to the role of the host sex and age, and hints to other avenues of research and strategies for their treatment and control.

[First cases of squamous cell carcinoma associated with cosmetic use of bleaching compounds]. / Premiers cas de carcinomes épidermoïdes sur terrain de dépigmentation artificielle.

INTRODUCTION: We report two cases of squamous cell carcinoma (SCC) in two black women (phenotype VI) using bleaching compounds for cosmetic purposes over a period of 15 years. CASE REPORTS: Two women (aged 45 and 47 years) with a long history of cosmetic use of bleaching compounds consulted at a dermatology unit for skin tumours. A diagnosis of SCC was confirmed by histological examination of tumour biopsies. One patient was HIV-positive. Surgical treatment was performed in both cases: simple postoperative complications were seen in one patient but the other died at home following recurrence of carcinoma in the year following diagnosis. DISCUSSION: To our knowledge, theses two cases represent the first description of SCC occurring after prolonged cosmetic use of bleaching compounds. Carcinoma occurred in both cases in skin exposed to sun. In our patients, the mechanism of carcinogenesis may have involved melanin destruction, solar exposure and corticosteroid-induced immunosuppression. A direct carcinogenic effect of hydroquinone or other unidentified compounds is another possibility; the carcinogenicity of hydroquinone is well established in rodents. While these observations do not provide formal proof of any implication of depigmentation products in SCC, they emphasize the need for monitoring of dark-skinned women using skin lighteners.

Progression of malaria induced pathogenicity during chloroquine therapy.

Treatment Failure with chloroquine is one of the challenges that faced the dedicated efforts to eradicate malaria This study aims at investigating the impact of treatment failure with chloroquine on the progression of the disease-induced histo-pathogenic and immunogenic outcomes. To achieve this, Rane's protocol with modifications was applied on a model of Plasmodium berghei ANKA infected ICR mice to determine the dose response curve of chloroquine and to screen the treatment impact on the disease progression. Chloroquine was given at 1, 5, 10, 15 and 20 mg/kg once the parasitemia reached to 20-30% (the experimental initiation point). During the subsequent days, the mice were monitored for changes in the clinical signs, hematology parameters and the progress of the parasitemia until the parasitemia reached to 60-70% (the experimental termination point) or up to 10 days after chloroquine administration in case of achieving a complete eradication of the parasite. At the end, the mice were exsanguinated and their blood and organs were collected for the biochemistry and the histology study. A complete eradication of the parasite was achieved at 20 mg/kg while recrudescence was observed at the lower doses. At 1 mg/kg, the parasite growth was comparable to that of the positive control. The histo-pathogenic and immunogenic changes were stronger in the groups that experienced recrudescence (at 5 and 10 mg/kg). All in all, the study highlights the possibility of having a worsened clinical condition when chloroquine is given at its sub-therapeutic doses during malaria treatment.

Characterization of a new genotype and serotype of infectious bronchitis virus in Western Africa.

Between 2002 and 2007, more than 1000 chickens from commercial farms, live bird markets and backyard farms in Nigeria and Niger were tested for the presence of the infectious bronchitis virus (IBV) genome. Phylogenetic analysis of full-length sequences of the spike 1 (S1) gene revealed a new genotype of IBV that we refer to as 'IBADAN'. The minimum genetic distance to the closest 'non-IBADAN' strains (UK/7/93 at the nucleotide level; H120 and M41 at the amino acid level) reached 24 and 32 % at the nucleotide and amino acid levels, respectively. The full genome of the IBADAN reference strain (NGA/A116E7/2006) had a genetic distance of 9.7-16.4 % at the nucleotide level with all available fully sequenced strains. As IBV S1 plays a major role in antigenicity, the antigenic relatedness of NGA/A116E7/2006 was compared with strains of other serotypes. NGA/A116E7/2006 did not cross-react with antisera against IT02, M41, D274, Connecticut or 793/B strains in virus neutralization assays. NGA/A116E7/2006 cross-reacted with the QX-like strain ITA/90254/2005 but only to a low level (antigenic relatedness of 33 %), suggesting that IBADAN also represents a new serotype. A comparison of S1 sequences identified several amino acids that may play a role in IBV antigenicity. Despite the absence of obvious clinical signs in poultry infected by IBADAN strains, it is important to test the cross-protection of current vaccine strains.

Newcastle disease virus in West Africa: new virulent strains identified in non-commercial farms.

Forty-four Newcastle disease virus (NDV) strains, obtained between 2002 and 2007 from different poultry species in Nigeria, Niger, Burkina Faso and Cameroon, were phylogenetically analysed based on partial F sequences. Lineage 2 viruses were genetically identical or similar to the locally used LaSota vaccine strain and were mostly detected in commercial farms. Lineage 1, 3 and 4 strains were only sporadically found, and their origin was less clear. Twenty-one strains from backyard farms and live bird markets formed three new clusters within lineage 5, tentatively named 5f, 5g and 5h. All of these strains were predicted to be virulent based on their F protein cleavage site sequence. Minimal genetic distances between new and previously established sublineages ranged from 9.4 to 15.9%, and minimal distances between the new sublineages were 11.5 to 17.3%. Their high genetic diversity and their presence in three different Sub-Saharan countries suggest that these new sublineages represent the NDV variants indigenous to West Africa.

Whole genome analysis of the action of interferon-beta.

OBJECTIVES: To characterize the IFNbeta1a-regulated gene expression on leukocytes of Multiple Sclerosis (MS) patients using microarrays with whole human genome representation. METHODS: Genes differentially expressed by interferon-beta were identified by a microarray in vitro study performed in leukocytes obtained from 5 MS relapsing-remitting patients. RESULTS: Following the culture of peripheral blood mononuclear cells from MS relapsing-remitting patients for 24 hs with IFNbeta1a, the expression of 868 genes was modified: 545 increased (including CXCL11, CCL8, INDO, IFI27, CFB, CXCL10 and IFIT1) and 323 diminished (including RBP7, SEPT5, RNF8, ADORA2B and FOS). CONCLUSIONS: Since many of them were previously recognized as involved in MS pathogenesis, the IFNbeta1a mechanism of action could imply a compensatory regulation of systems deregulated in MS.

(-)­Oleocanthal inhibits proliferation and migration by modulating Ca2+ entry through TRPC6 in breast cancer cells.

Triple negative breast cancer is an aggressive type of cancer that does not respond to hormonal therapy and current therapeutic strategies are accompanied by side effects due to cytotoxic actions on normal tissues. Therefore, there is a need for the identification of anti-cancer compounds with negligible effects on non-tumoral cells. Here we show that (-)­oleocanthal (OLCT), a phenolic compound isolated from olive oil, selectively impairs MDA-MB-231 cell proliferation and viability without affecting the ability of non-tumoral MCF10A cells to proliferate or their viability. Similarly, OLCT selectively impairs the ability of MDA-MB-231 cells to migrate while the ability of MCF10A to migrate was unaffected. The effect of OLCT was not exclusive for triple negative breast cancer cells as we found that OLCT also attenuate cell viability and proliferation of MCF7 cells. Our results indicate that OLCT is unable to induce Ca2+ mobilization in non-tumoral cells. By contrast, OLCT induces Ca2+ entry in MCF7 and MDA-MB-231 cells, which is impaired by TRPC6 expression silencing. We have found that MDA-MB-231 and MCF7 cells overexpress the channel TRPC6 as compared to non-tumoral MCF10A and treatment with OLCT for 24-72 h downregulates TRPC6 expression in MDA-MB-231 cells. These findings indicate that OLCT impairs the ability of breast cancer cells to proliferate and migrate via downregulation of TRPC6 channel expression while having no effect on the biology of non-tumoral breast cells.

Two recombinant human interferon-beta 1a pharmaceutical preparations produce a similar transcriptional response determined using whole genome microarray analysis.

OBJECTIVES: Recombinant human interferon-beta (IFN-b) is a well-established treatment for multiple sclerosis (MS). The regulatory process for marketing authorization of biosimilars is currently under debate in certain countries. In the EU, EMEA has clearly defined the process including overarching and product-specific guidelines, which includes clinical testing. Biosimilarity needs to be based on comparability criteria, including at least molecular characterization, biological activity relevant for the therapeutic effect and relative bioavailability ("bioequivalence"). In the case of such complex diseases as MS, where the effect of treatment is not so directly measurable, in vitro tools can provide additional data to support comparability. Genomic microarrays assays might be useful to compare multisource biopharmaceuticals. The aim of the present study was to compare the pharmacodynamic genomic effects (in terms of transcriptional regulation) of two recombinant human IFN-I(2)1a preparations on lymphocytes of multiple sclerosis patients using a whole genome microarray assay. METHODS: We performed an ex vivo whole genome expression profiling of the effect of two preparations of IFN-I(2)1a on non-adherent mononuclears from five relapsing-remitting MS patients analyzing microarrays (CodeLink Human Whole Genome). Patients blood was drawn, PBMCs isolated and cultured in three different conditions: culture medium (control), 1,000 U/ml of IFN-I(2)1a (BLA- (STOFERON, Bio Sidus) and 1,000 U/ml of IFN-I(2)1a (REBIF, Serono) RNA was purified from non-adherent cells (mostly lymphocytes), amplified and hybridized. Raw data were generated by CodeLink proprietary software. Data normalization, quality control and analysis of differential gene expression between treatments were done using linear model for microarray data. Functional annotation analysis of IFN-I(2)1a MS treatment transcription was done using DAVID. RESULTS: Out of the approximately 45,000 human sequences examined, no evidence of differential regulation was found when both treatments were compared (minimum adjusted p-value > 0.999). The IFN-I(2)1a effect differentially regulated the expression of 868 genes. The expression of standard markers such as GTP cyclohidrolase, MxA, and OAS isoenzymes A and B changed as a consequence of the action of IFN-I(2)1a. CONCLUSIONS: This exhaustive and highly sensitive assay did not show differences in the genomic expression profile of these two products under the assayed experimental conditions. These results suggest that this technology might be useful for the initial comparison of biosimilars, being part of a comprehensive comparability program that includes clinical testing.