Multiplex diagnostics : development of serological dualmode multiplex microarray immunoassay and analysis of influenza and respiratory syncytial virus disease burden in early childhood

Respiratory syncytial virus (RSV) and influenza A (IAV) and B (IBV) viruses infect humans throughout their life with exceptionally high rates of re-infection. Serological assays are commonly used to diagnose and characterize host immune responses against microbial pathogens acquired by natural infection or vaccination. The current methods are mostly based on enzyme immunoassay (EIA) or other conventional methods in a single analyte format. Presently, there is a need to shift from single analytical methods to microarray format which enables the detection of antibodies against multiple targets in a rapid and cost-effective manner. In this study, a highly sensitive single analyte EIA method was used for serological analyses in children’s sera. In addition, multiplex microarray immunoassay (MAIA) methods were developed for rapid and simultaneous detection of IgG antibodies against seven viral antigens (H1N1pdm09 vaccine ag, IAV H1N1, IAV H3N2, IBV Victoria, IBV Yamagata, RSV and adenovirus hexon protein). We found out that MAIA is well suitable for large-scale serosurveillance and vaccine immunity studies. We followed-up virus-specific immunity by MAIA in response to natural infection and vaccination in a large cohort of 0-2 year old children. Our serological findings showed a high rate of respiratory virus infections and reinfections in young children. The applicability of MAIA in vaccine immunity studies was also analysed. We developed a specific, sensitive, sample and antigen saving assay for simultaneous detection of IgM and IgG antibodies against H1N1pdm09 vaccine ag and IBV Yamagata. MAIA showed excellent correlation with EIA and a good correlation with hemagglutination inhibition assay in measurement of vaccine-induced antibodies in sera of Pandemrix-vaccinated adults.Respiratory syncytial virus (RSV) ja influenssa A- (IAV) ja B(IBV)-virukset ovat merkittäviä ylähengitystieinfektioiden aiheuttajia koko ihmisen elinkaaren ajan. RSV:n ja influenssan aiheuttamat infektiot voivat joskus olla vakavia tai jopa henkeä uhkaavia erityisesti pienillä lapsilla ja ikäihmisillä. Nykyiset serodiagnostiset menetelmät perustuvat entsyymi-immunologisiin (EIA) ja muihin määritysmenetelmiin, joissa immuunivastetta voidaan tutkia kerrallaan vain yhtä taudinaiheuttajaa kohtaa. Tällä hetkellä on seerumin vasta-ainetutkimusten osalta suuri tarve siirtyä monianalyyttisiin menetelmiin, jotka olisivat nopeampia ja kustannustehokkaampia kuin nykyiset tutkimusmenetelmät. Tässä tutkimuksessa on käytetty herkkiä EIA-menetelmiä pienten lasten ja influenssarokotettujen henkilöiden seerumien virusspesifisen immuunivasteen tutkimiseen. Työssä on myös kehitetty monianalyyttinen mikrosiruperusteinen immunomääritysmenetelmä (MAIA) samanaikaiseen seerumin IgG-luokan vastaaineiden mittaamiseen eri influenssavirusantigeeneja, RSV:tä ja adenoviruksen heksoniproteiinia kohtaan. Tutkimuksemme osoittivat, että hengitysteiden virusinfektiot ja yllämainittujen virusten aiheuttamat uusintainfektiot ovat erittäin tavallisia pienillä lapsilla. MAIA-menetelmää sovellettiin myös aikuisten influenssarokotevasteiden analyysiin. Menetelmää kehitettiin edelleen siten, että näytteestä voitiin samalla kertaa mitata seerumin IgG- ja IgM-luokan vasta-aineita eri IAV- ja IBV-virusantigeeneja kohtaan. Menetelmä osoittautui erittäin spesifiseksi, herkäksi ja näytettä ja antigeeneja säästäväksi ja tulokset olivat hyvin yhtenevät muiden perinteisten vasta-ainetutkimusmenetelmien kanssa. Kehittämämme MAIA-menetelmä toimi erittäin hyvin ja sen voitiin osoittaa sopivan erinomaisesti laajojen väestön seerumiaineistojen ja virusrokotevasteiden analyysiin

PFKFB3 overexpression in monocytes of patients with colon but not rectal cancer programs pro-tumor macrophages and is indicative for higher risk of tumor relapse

Introduction: Circulating monocytes are main source for tumor-associated macrophages (TAMs) that control tumor growth, angiogenesis, metastasis and therapy resistance. We raised the questions how monocyte programming is affected by growing tumors localized in colon and rectal sections, and how treatment onsets affect monocyte programming in the circulation. Methods: Patients with rectal cancer and colon cancer were enrolled in the study. Peripheral blood monocytes were characterized by phenotypic analysis using flow cytometry, by transcriptomic analysis using RNA sequencing and by gene expression analysis using real-time RT-PCR. Phenotypic analysis was performed with IF/confocal microscopy. Spatial transcriptomic analysis was applied using GeoMX DSP-NGS. Results: In patients with rectal cancer, increased amount of CCR2+ monocytes was indicative for the absence of both lymphatic and hematogenous metastasis. In contrast, in patients with colon cancer CD163+ monocytes were indicative for LN metastasis. NGS analysis identified tumor-specific transcriptional programming of monocytes in all CRC patients compared to healthy individuals. The key transcriptional difference between monocytes of patients with colon and rectal cancer was increased expression of PFKFB3, activator of glycolysis that is currently considered as therapy target for major solid cancers. PFKFB3-expressing monocyte-derived macrophages massively infiltrated tumor in colon. Nanostring technology identified correlation of PFKFB3 with amount and tumor-promoting properties of TAMs in colon but not in rectal cancer. PFKFB3 was indicative for tumor relapse specifically in colon cancer. Discussion: Our findings provide essential argument towards CRC definition to cover two clinically distinct cancers – colon cancer and rectal cancer, that differentially interact with innate immunity

Factors influencing the sexual drive of Russian women of reproductive age in the digital age

Aim. To assess the impact of age, number of sexual partners, having children, family income, and time spent with a smartphone or computer on the sexual drive of women of reproductive age in the era of information technology. Materials and methods. A study of the sexual drive of 79 women aged 18–35 in family (partner) relationships was conducted. The well-established international Female Sexual Function Index was used. Respondents also answered questions about some aspects of personality and social status. The results were evaluated using the methods of mathematical statistics: descriptive statistics, correlation, determination, and logit-regression analyses. Results. The intensity of sexual drive correlates with a woman's age, the number of sexual partners, and the family income. There was no correlation between the level of a woman's sexual drive and having children and the time women and their partners spend with computers and smartphones

(E)-17β,19-Epoxy­methano-17,23,24-tridemethyl-4-nor-5β,18α-olean-3-one oxime

In the penta­cyclic triterpenoide skeleton of the title mol­ecule, C27H43NO2 [systematic name: (3E,3aS,5aR,5bR,7aR,11R,11aR,11bR,13aR,13bR)-5a,5b,10,10,13b-penta­methyl­icosa­hydro-1H-11,7a-(epoxy­methano)cyclo­penta­[a]chrysen-3-one oxime], the five-membered ring A has an envelope conformation, while the six-membered rings B–E adopt chair conformations. Rings A and B are cis-fused. The hydroximino group has an E configuration. Strong inter­molecular O—H⋯O hydrogen bonds link the mol­ecules into helical chains

Experience of inulin use for correcting intestinal microbiota in patients suffering from recurrent vulvovaginal candidiasis: A prospective cohort comparative study

Aim. To evaluate the effectiveness of inulin for the adjustment of intestinal microflora in patients with recurrent vulvovaginal candidiasis (VVC). Materials and methods. A prospective cohort comparative study included 79 women aged 18 to 50 years. They were divided into three groups: the main group included 32 patients receiving complex treatment with a dietary supplement containing inulin derived from the "Extra" variety of Jerusalem artichoke root in combination with fluconazole; the comparison group included 27 patients receiving fluconazole only; and the control group included 20 healthy women. The patients were followed up for 12 months. Examination included stool culture for dysbiosis, complete blood count, urinalysis, blood chemistry (glucose, bilirubin, alanine aminotransferase, aspartate aminotransferase, total protein, urea, creatinine), examination of the vulva and vaginal mucosa, vaginal smear microscopy for microflora, Gram staining, real-time polymerase chain reaction for Mycoplasma hominis, Ureaplasma parvum, Trihomonas vaginalis, Chlamydia traсhomatis, Candida albicans. Subsequently, all subjects every three months underwent a bimanual examination, speculum examination of the cervical mucosa and vagina, Gram staining of vaginal mucosa and cervix swabs, a thorough interview, and history taking. At 12 months, stool was cultured for dysbiosis again, and the data were analyzed. Results. The VVC therapy in the group of inulin-containing dietary supplement was more effective: the recurrence rate was 3 times lower than with the standard treatment regimen. Conclusion. The use of dietary supplement containing inulin derived from the "Extra" variety of Jerusalem artichoke root in complex therapy for recurrent VVC can significantly improve the gut and vagina microflora condition and prevent the recurrence and normalization of stool in patients

PFKFB3 overexpression in monocytes of patients with colon but not rectal cancer programs pro-tumor macrophages and is indicative for higher risk of tumor relapse

IntroductionCirculating monocytes are main source for tumor-associated macrophages (TAMs) that control tumor growth, angiogenesis, metastasis and therapy resistance. We raised the questions how monocyte programming is affected by growing tumors localized in colon and rectal sections, and how treatment onsets affect monocyte programming in the circulation.MethodsPatients with rectal cancer and colon cancer were enrolled in the study. Peripheral blood monocytes were characterized by phenotypic analysis using flow cytometry, by transcriptomic analysis using RNA sequencing and by gene expression analysis using real-time RT-PCR. Phenotypic analysis was performed with IF/confocal microscopy. Spatial transcriptomic analysis was applied using GeoMX DSP-NGS.ResultsIn patients with rectal cancer, increased amount of CCR2+ monocytes was indicative for the absence of both lymphatic and hematogenous metastasis. In contrast, in patients with colon cancer CD163+ monocytes were indicative for LN metastasis. NGS analysis identified tumor-specific transcriptional programming of monocytes in all CRC patients compared to healthy individuals. The key transcriptional difference between monocytes of patients with colon and rectal cancer was increased expression of PFKFB3, activator of glycolysis that is currently considered as therapy target for major solid cancers. PFKFB3-expressing monocyte-derived macrophages massively infiltrated tumor in colon. Nanostring technology identified correlation of PFKFB3 with amount and tumor-promoting properties of TAMs in colon but not in rectal cancer. PFKFB3 was indicative for tumor relapse specifically in colon cancer.DiscussionOur findings provide essential argument towards CRC definition to cover two clinically distinct cancers – colon cancer and rectal cancer, that differentially interact with innate immunity

t(11;16)(q23;q24) KMT2A/USP10

Review on t(11;16)(q23;q24), with data on clinics, and the genes involved

Results of the COVID-19 mental health international for the general population (COMET-G) study.

INTRODUCTION: There are few published empirical data on the effects of COVID-19 on mental health, and until now, there is no large international study. MATERIAL AND METHODS: During the COVID-19 pandemic, an online questionnaire gathered data from 55,589 participants from 40 countries (64.85% females aged 35.80 ± 13.61; 34.05% males aged 34.90±13.29 and 1.10% other aged 31.64±13.15). Distress and probable depression were identified with the use of a previously developed cut-off and algorithm respectively. STATISTICAL ANALYSIS: Descriptive statistics were calculated. Chi-square tests, multiple forward stepwise linear regression analyses and Factorial Analysis of Variance (ANOVA) tested relations among variables. RESULTS: Probable depression was detected in 17.80% and distress in 16.71%. A significant percentage reported a deterioration in mental state, family dynamics and everyday lifestyle. Persons with a history of mental disorders had higher rates of current depression (31.82% vs. 13.07%). At least half of participants were accepting (at least to a moderate degree) a non-bizarre conspiracy. The highest Relative Risk (RR) to develop depression was associated with history of Bipolar disorder and self-harm/attempts (RR = 5.88). Suicidality was not increased in persons without a history of any mental disorder. Based on these results a model was developed. CONCLUSIONS: The final model revealed multiple vulnerabilities and an interplay leading from simple anxiety to probable depression and suicidality through distress. This could be of practical utility since many of these factors are modifiable. Future research and interventions should specifically focus on them