Human papillomavirus genotypes in male genitalia and their concordance among pregnant spouses participating in the Finnish Family HPV study

IntroductionGenotype‐specific concordance among human papillomavirus (HPV)‐infected spouses is incompletely assessed. HPV concordance is of importance in counselling HPV‐infected subjects and their partners.AimTo determine HPV‐genotype distribution and prevalence in semen and urethral samples of young fathers and their concordance with HPV‐genotypes in cervical and oral mucosal samples of their spouses.MethodsIn the Finnish Family HPV study, urethral and semen samples were collected from 131 fathers‐to‐be for HPV genotyping with Multimetrix assay. HPV‐genotyping data were correlated with the data of the cervical and oral mucosal samples of the spouses collected at third trimester pregnancy at baseline.Main Outcome MeasureConcordance of HPV‐genotypes between the spouses.ResultsUrethral and/or semen samples tested HPV positive in 47/131 (35.9%) of the fathers‐to‐be, more frequently in semen (28/90; 31%) than in urethral (29/128; 23%) samples. In women, 18.8% of their cervical and 17.2% of the oral samples tested HPV‐positive. Of the HPV‐positive samples, HPV16 was the most frequent genotype, present in 39.3% of semen, 77.3% of mothers' oral, and 29.2% of cervical samples. Multiple‐type infections were found in 24.1% of HPV‐positive urethral and 21.6% of semen samples. In mothers, half of their cervical and 4.5% of oral HPV were multiple‐type infections. The HPV‐genotype‐specific concordance among spouses ranged from 0% to 9.5%, according to the sampling sites. Altogether, eight couples disclosing such a concordance were analyzed separately for a risk‐profile. Mothers of the HPV‐concordant couples reported significantly higher number (>6) of life‐time sexual partners than did the discordant mothers (P = 0.030).ConclusionsAsymptomatic HPV infections were common in both parents, while the genotype‐specific concordance among spouses was low. In both partners, HPV16 is the dominant genotype. HPV6 was a prevalent HPV genotype in male genitalia. Female risk profile might enhance the transmission of HPV infection to her male partner and increase the genotype‐specific HPV concordance between the spouses. Kero K, Rautava J, Syrjänen K, Grenman S, and Syrjänen S. Human papillomavirus genotypes in male genitalia and their concordance among pregnant spouses participating in the Finnish family HPV study. J Sex Med 2011;8:2522–2531.     

Long-term exposure to intermittent hypoxia results in increased hemoglobin mass,reduced plasma volume,and elevated erythropoietin plasma levels in man

While it is well established that highlanders have optimized their oxygen transport system, little is known about the acclimatization of those who move between different altitudes. The purpose of this study was to establish whether the acclimatization to long-term intermittent hypoxic exposure in members of the Chilean Army who frequently move from sea level to 3,550 m altitude is correlated with acute acclimatization or chronic adaptation to hypoxia. A group of officers was exposed intermittently to hypoxia for about 22 years (OI, officers at intermittent hypoxia) and a group of soldiers for 6 months (SI, soldiers at intermittent hypoxia). Both groups were compared to residents at altitude (RA) and to soldiers at sea level (SL). When compared to SL, we observed an 11% increase in total hemoglobin mass (tHb) as well as a corresponding increase in red cell volume (RCV), hemoglobin concentration and hematocrit in all three groups at altitude. Plasma volume (PV) and blood volume (BV) decreased at altitude but increased when OI and SI returned to sea level. Moreover, intermittent hypoxic exposure of OI and SI resulted in increased plasma erythropoietin (Epo) levels, which peaked on day 2 at high altitude followed by decreasing levels during the successive days, and reaching pre-altitude values in SI even when staying at altitude. In conclusion, with regard to tHb and RCV, the acclimatization to long-term intermittent hypoxia resembles the adaptation to chronic hypoxia, while PV and BV regulation mimicked acclimatization to acute hypoxia. Remarkably, finely controlled regulation of Epo expression still occurs after up to 22 years of weekly exposure to altitude. Electronic Publication     

Recombinant human beta 2-defensin fusion proteins as a tool to investigate defensin structure and function in small human intestinal tissue samples

Objective

Effects of immune cells on the beta 2 (β2)-defensin (HBD2) expression and its antibacterial activity in the intestinal mucosa of patients with inflammatory bowel diseases remains unclear. The small size of these proteins presents a major challenge in localizing antibacterial activities in human intestinal tissue. In this study, we evaluated the detection limits at mRNA and protein level by approaching HBD2 from small tissue samples.

Methods

HT-29 colonic epithelial cells were incubated with proinflammatory cytokines before HBD2 mRNA was investigated by quantitative polymerase chain reaction. The HBD2 protein was assessed by Western blot analysis using HBD2 fused with enhanced green fluorescent protein (HBD2-EGFP). Purified HBD2 fused with the glutathione-S-transferase (GST-HBD2) was used to detect antibacterial activity in a densitometric assay.

Results

Interleukin (IL)-1β induced HBD2 mRNA in HT-29 cells; however, tumor necrosis factor-α, IL-6 and IL-17 did not. The Western blot had a sensitivity of 1.5?pmol to detect recombinant HBD2, but did not detect HBD2 in either human intestinal or IL-1β-treated HT-29 cells. HBD2-EGFP was detected by HBD2-specific Western blot within cell lysates and culture supernants of transfected HT-29 and primary cells. In nanomolar ranges, GST-HBD2 reduced bacterial growth. The HBD2 bioactivity depended on solution conditions, but not on the size of the fusion partner.

Conclusion

The established fusion proteins provide excellent tools to evaluate expression patterns and antibacterial effects of HBD2 in human intestinal tissue samples.     

ALDH1A1-related stemness in high-grade serous ovarian cancer is a negative prognostic indicator but potentially targetable by EGFR/mTOR-PI3K/aurora kinase inhibitors

Poor chemotherapy response remains a major treatment challenge for high-grade serous ovarian cancer (HGSC). Cancer stem cells are the major contributors to relapse and treatment failure as they can survive conventional therapy. Our objectives were to characterise stemness features in primary patient-derived cell lines, correlate stemness markers with clinical outcome and test the response of our cells to both conventional and exploratory drugs. Tissue and ascites samples, treatment-naive and/or after neoadjuvant chemotherapy, were prospectively collected. Primary cancer cells, cultured under conditions favouring either adherent or spheroid growth, were tested for stemness markers; the same markers were analysed in tissue and correlated with chemotherapy response and survival. Drug sensitivity and resistance testing was performed with 306 oncology compounds. Spheroid growth condition HGSC cells showed increased stemness marker expression (including aldehyde dehydrogenase isoform I; ALDH1A1) as compared with adherent growth condition cells, and increased resistance to platinum and taxane. A set of eight stemness markers separated treatment-naive tumours into two clusters and identified a distinct subgroup of HGSC with enriched stemness features. Expression of ALDH1A1, but not most other stemness markers, was increased after neoadjuvant chemotherapy and its expression in treatment-naive tumours correlated with chemoresistance and reduced survival. In drug sensitivity and resistance testing, five compounds, including two PI3K-mTOR inhibitors, demonstrated significant activity in both cell culture conditions. Thirteen compounds, including EGFR, PI3K-mTOR and aurora kinase inhibitors, were more toxic to spheroid cells than adherent cells. Our results identify stemness markers in HGSC that are associated with a decreased response to conventional chemotherapy and reduced survival if expressed by treatment-naive tumours. EGFR, mTOR-PI3K and aurora kinase inhibitors are candidates for targeting this cell population. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Differential diagnosis of vacuolar myopathies in the NGS era

Altered autophagy accompanied by abnormal autophagic (rimmed) vacuoles detectable by light and electron microscopy is a common denominator of many familial and sporadic non‐inflammatory muscle diseases. Even in the era of next generation sequencing (NGS), late‐onset vacuolar myopathies remain a diagnostic challenge. We identified 32 adult vacuolar myopathy patients from 30 unrelated families, studied their clinical, histopathological and ultrastructural characteristics and performed genetic testing in index patients and relatives using Sanger sequencing and NGS including whole exome sequencing (WES). We established a molecular genetic diagnosis in 17 patients. Pathogenic mutations were found in genes typically linked to vacuolar myopathy (GNE, LDB3/ZASP, MYOT, DES and GAA), but also in genes not regularly associated with severely altered autophagy (FKRP, DYSF, CAV3, COL6A2, GYG1 and TRIM32) and in the digenic facioscapulohumeral muscular dystrophy 2. Characteristic histopathological features including distinct patterns of myofibrillar disarray and evidence of exocytosis proved to be helpful to distinguish causes of vacuolar myopathies. Biopsy validated the pathogenicity of the novel mutations p.(Phe55*) and p.(Arg216*) in GYG1 and of the p.(Leu156Pro) TRIM32 mutation combined with compound heterozygous deletion of exon 2 of TRIM32 and expanded the phenotype of Ala93Thr‐caveolinopathy and of limb‐girdle muscular dystrophy 2i caused by FKRP mutation. In 15 patients no causal variants were detected by Sanger sequencing and NGS panel analysis. In 12 of these cases, WES was performed, but did not yield any definite mutation or likely candidate gene. In one of these patients with a family history of muscle weakness, the vacuolar myopathy was eventually linked to chloroquine therapy. Our study illustrates the wide phenotypic and genotypic heterogeneity of vacuolar myopathies and validates the role of histopathology in assessing the pathogenicity of novel mutations detected by NGS. In a sizable portion of vacuolar myopathy cases, it remains to be shown whether the cause is hereditary or degenerative.     

Maternal anxiety disorders prior to conception, psychopathology during pregnancy and early infants’ development: a prospective-longitudinal study

Family-genetic studies suggest that anxiety disorders run in families and that mechanisms of familial transmission might act as early as during pregnancy. The aims of the Maternal Anxiety in Relation to Infant Development (MARI) Study are to prospectively investigate the course of pregnancy in women with and without anxiety disorders prior to conception from early pregnancy to postpartum focussing on (a) maternal psychopathology, (b) maternal perinatal health, and (c) offspring outcomes that are supposed to be early indicators/ antecendents for later anxiety disorders. The MARI Study is a prospective-longitudinal study program with seven waves of assessment: T1 (baseline: week 10 to 12 of gestation), T2 (week 22 to 24 of gestation), T3 (week 35 to 37 of gestation), T4 (10 days postpartum), T5 (2 months postpartum), T6 (4 months postpartum), and T7 (16 months postpartum). Overall, N?=?306 pregnant women were enrolled during early pregnancy (T1) and allocated to one of the following initial diagnostic groups: no AD: no anxiety nor depressive disorder prior to pregnancy (N?=?109), pure D: pure depressive disorder(s) prior to pregnancy (N?=?48), pure A: pure anxiety disorder(s) prior to pregnancy (N?=?84), and comorbid AD: comorbid anxiety and depressive disorders prior to pregnancy (N?=?65). Overall, N?=?284 mothers could be retained until T6 (retention rate: 92.8 %) and N?=?274 until T7 (retention rate: 89.5 %). Clinical and psychosocial measures were used including a standardized diagnostic interview (CIDI-V) with dimensional scales and standardized observation paradigms (mother-infant-relationship, infant temperament and neuropsychological development). Dimensional anxiety and depression liability indices were developed to reflect the severity of anxiety and depressive disorders prior to pregnancy and to ease longitudinal modelling. Findings from this study will contribute to improved knowledge about the natural course of anxiety disorders during transition to parenthood and associated outcomes that are assumed to be early indicators of later psychopathology in the offspring. Results are expected to provide new insights into mechanisms of familial transmission and clues for targeted prevention and early intervention.     

T‐bet is essential for Th1‐mediated,but not Th17‐mediated,CNS autoimmune disease

T cells that produce both IL‐17 and IFN‐γ, and co‐express ROR‐γt and T‐bet, are often found at sites of autoimmune inflammation. However, it is unknown whether this co‐expression of T‐bet with ROR‐γt is a prerequisite for immunopathology. We show here that T‐bet is not required for the development of Th17‐driven experimental autoimmune encephalomyelitis (EAE). The disease was not impaired in T‐bet^?/? mice and was associated with low IFN‐γ production and elevated IL‐17 production among central nervous system (CNS) infiltrating CD4⁺ T cells. T‐bet^?/? Th17 cells generated in the presence of IL‐6/TGF‐β/IL‐1 and IL‐23 produced GM‐CSF and high levels of IL‐17 and induced disease upon transfer to naïve mice. Unlike their WT counterparts, these T‐bet^?/– Th17 cells did not exhibit an IL‐17→IFN‐γ switch upon reencounter with antigen in the CNS, indicating that this functional change is not critical to disease development. In contrast, T‐bet was absolutely required for the pathogenicity of myelin‐responsive Th1 cells. T‐bet‐deficient Th1 cells failed to accumulate in the CNS upon transfer, despite being able to produce GM‐CSF. Therefore, T‐bet is essential for establishing Th1‐mediated inflammation but is not required to drive IL‐23‐induced GM‐CSF production, or Th17‐mediated autoimmune inflammation.     

Examining the validity of the unitary theory of clinical relationships: comparison of observed and experienced parent-doctor interaction

Objective

We explored parent–doctor relationships in the care of children with leukaemia from three perspectives simultaneously: parents’, doctors’ and observers’. Our aim was to investigate convergence and divergence between these perspectives and thereby examine the validity of unitary theory of emotionality and authority in clinical relationships.

Methods

33 audiorecorded parent–doctor consultations and separate interviews with parents and doctors, which we analysed qualitatively and from which we selected three prototype cases.

Results

Across the whole sample doctors’ sense of relationship generally converged with our observations of consultation, but parents’ sense of relationship diverged strongly from each. Contrary to current assumptions, parents’ sense of emotional connection with doctors did not depend on doctors’ emotional behaviour, and parents did not feel disempowered by doctors’ authority. Moreover, authority and emotionality were not conceptually distinct for parents, who gained emotional support from doctors’ exercise of authority.

Conclusions

The relationships looked very different from the three perspectives. These divergences indicate weaknesses in current ideas of emotionality and authority in clinical relationships and the necessity of multisource datasets to develop these ideas in a way that characterises clinical relationships from all perspectives.

Practice implications

Methodological development will be needed to address the challenges posed by multisource datasets.     

Antibodies against acute phase proteins and their functions in the pathogenesis of disease: a collective profile of 25 different antibodies

The acute phase response is a defense system in which the innate immune response is activated following injury or infection. Positive and negative acute phase proteins (APPs) are crucial for protecting the host organism, as well as returning it to homeostatic levels, the first with elevated concentrations and the latter with decreased concentrations during the acute phase. Reports about the presence of antibodies against APPs are known, however their individual, as well as potentially collective, pathological or physiological roles are still emerging. Some of these autoantibodies are specifically connected with diseases (such as pancreatic secretory trypsin inhibitor and C3, C4 nephritic factors), while others have been reported as natural antibodies. The persistent presence (even if only minor) of autoantibodies in healthy blood donors indicates an overlapping category of autoantibodies, which could become pathogenic, depending on the autoantibody characteristics such as avidity, epitope specificity, changes in the microenvironment leading to different oxidative status and others. This review uses the novel approach of studying the overall autoantibody population against APPs, their functions and connections to diseases. The primary function of autoantibodies against APPs (anti-APPs) is thought to promote their clearance, however autoantibodies against negative APPs have also been found and applying the same role to those is doubtful. There is also the theory of consumption in the stage of inflammation, which could be relevant to anti-APPs. Reports about protective roles of autoantibodies are also emerging, showing lowered levels of antibodies in diseases, which could be interesting for therapeutic intervention.     

Suppression of IgE-mediated anaphylaxis and food allergy with monovalent anti-FcεRIα mAbs

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