Temporal variation in climatic factors influences phenotypic diversity of Trochulus land snails.

Organisms with limited dispersal capabilities should show phenotypic plasticity in situ to keep pace with environmental changes. Therefore, to study the influence of environmental variation on the phenotypic diversity, we chose land snails, Trochulus hispidus and T. sericeus, characterized by high population variability. We performed long-term field studies as well as laboratory and common garden experiments, which revealed that temporal environmental changes generate visible variation in shell size and shape of these snails. Many shell measurements of T. hispidus varied significantly with temperature and humidity in individual years. According to this, the first generation of T. hispidus, bred in controlled laboratory conditions, became significantly different in higher spire and narrower umbilicus from its wild parents. Interestingly, offspring produced by this generation and transplanted to wild conditions returned to the 'wild' flat and wide-umbilicated shell shape. Moreover, initially different species T. hispidus and T. sericeus transferred into common environment conditions revealed rapid and convergent shell modifications within one generation. Such morphological flexibility and high genetic variation can be evolutionarily favored, when the environment is heterogeneous in time. The impact of climate change on the shell morphometry can lead to incorrect taxonomic classification or delimitation of artificial taxa in land snails. These findings have also important implications in the context of changing climate and environment.

Diabetic ketoacidosis incidence among children with new-onset type 1 diabetes in Poland and its association with COVID-19 outbreak-Two-year cross-sectional national observation by PolPeDiab Study Group.

BACKGROUND: There are several observations that the onset of coronavirus 19 (COVID-19) pandemic was associated with an increase in the incidence of diabetic ketoacidosis (DKA). However, due to heterogeneity in study designs and country-specific healthcare policies, more national-level evidence is needed to provide generalizable conclusions. OBJECTIVE: To compare the rate of DKA in Polish children diagnosed with type 1 diabetes (T1D) between the first year of COVID-19 pandemic (15 March 2020 to 15 March 2021) and the preceding year (15 March 2019 to 15 March 2020). METHODS: Reference centers in 13 regions (covering ~88% of Polish children) retrospectively reported all new-onset T1D cases in children from assessed periods, including DKA status at admission, administered procedures and outcomes. Secondly, we collected regions' demographic characteristics and the daily-reported number of COVID-19-related deaths in each region. RESULTS: We recorded 3062 cases of new-onset T1D (53.3% boys, mean age 9.5 ± 4.3 years old) of which 1347 (44%) had DKA. Comparing pre- and post-COVID-19 period, we observed a significant increase in the rate of DKA (37.5%-49.4%, p < .0001). The fraction of moderate (+5.4%) and severe (+3.4%) DKA cases increased significantly (p = .0089), and more episodes required assisted ventilation (+2.1%, p = .0337). Two episodes of DKA during 2020/2021 period were fatal. By region, change in DKA frequency correlated with initial COVID-19 death toll (March/April 2020) (R = .6, p = .0287) and change in T1D incidence (R = .7, p = .0080). CONCLUSIONS: The clinical picture of new-onset children T1D in Poland deteriorated over a 2-year period. The observed increase in the frequency of DKA and its severity were significantly associated with the overlapping timing of the COVID-19 epidemic.

Ongoing Speciation and Gene Flow between Taxonomically Challenging Trochulus Species Complex (Gastropoda: Hygromiidae).

Geographical isolation, selection and genetic drift can cause the geographical diversification of populations and lead to speciation. Land snail species in the genus Trochulus show overlaps in geographical ranges as well as in morphology, but genetic data do not always support the species-level taxonomy based on morphological characters. Such a group offers an excellent opportunity to explore the processes involved. We have addressed the problem by determining the status of the restricted endemic T. graminicola within the larger context of Trochulus taxonomy. We used an integrated approach based on morphological features, ecological preferences and two molecular markers: mitochondrial COI sequences and microsatellites. Comparison of these results demonstrated: (i) conchological distinction of T. striolatus and T. sericeus; (ii) anatomical, ecological and genetic differentiation of T. graminicola and (iii) concordance between morphological characters and mtDNA markers in T. striolatus. Moreover, our data showed an intricate evolutionary history within the genus Trochulus, which can be best explained by: (i) recent or ongoing gene flow between taxa or (ii) their large ancestral polymorphism. Both of these hypotheses suggest that diversification within this group of snails has occurred relatively recently. The mismatches between species defined on morphology and on molecular genetics indicate the complexity of the processes involved in the diversification of this genus.

Phenotypic plasticity can explain evolution of sympatric polymorphism in the hairy snail Trochulus hispidus (Linnaeus, 1758).

Morphological variation of snails from the genus Trochulus is so huge that their taxonomy is unclear. The greatest variability concerns forms hispidus and sericeus/plebeius, which are often considered as separate species. To evidence the species barriers, we carried out crossbreeding experiments between these two sympatric morphs. Moreover, we compared the shell morphology of laboratory-bred offspring with their wild parents to test if the variation can be explained by the phenotypic plasticity model. We found that the two Trochulus morphs show no reproductive barriers. The fecundity rates, the mean clutch size, and F1 viability observed for all crosses were not significantly different. In hybrid crosses (in F2 generation), we also recorded reproduction compatibility, similar fecundity, and hatching success as in their parents. Accordingly, phylogenetic analyses revealed the significant grouping of sequences from these different morphs and supported no constrains in reproduction between them. Comparison of shell morphology between wild and laboratory samples showed that various characters appeared highly plastic. The average shell shape of the hispidus morph changed significantly from flat with wide umbilicus to elevated with narrower umbilicus such as in the sericeus/plebeius morph. All these findings indicate that the examined morphs do not represent separate biological species and the evolutionary process is not advanced enough to separate their genetic pool. Therefore, phenotypic plasticity has played a significant role in the evolution of Trochulus shell polymorphism. The two morphs can evolve independently in separate phylogenetic lineages under the influence of local environmental conditions.

NONO couples the circadian clock to the cell cycle.

Mammalian circadian clocks restrict cell proliferation to defined time windows, but the mechanism and consequences of this interrelationship are not fully understood. Previously we identified the multifunctional nuclear protein NONO as a partner of circadian PERIOD (PER) proteins. Here we show that it also conveys circadian gating to the cell cycle, a connection surprisingly important for wound healing in mice. Specifically, although fibroblasts from NONO-deficient mice showed approximately normal circadian cycles, they displayed elevated cell doubling and lower cellular senescence. At a molecular level, NONO bound to the p16-Ink4A cell cycle checkpoint gene and potentiated its circadian activation in a PER protein-dependent fashion. Loss of either NONO or PER abolished this activation and circadian expression of p16-Ink4A and eliminated circadian cell cycle gating. In vivo, lack of NONO resulted in defective wound repair. Because wound healing defects were also seen in multiple circadian clock-deficient mouse lines, our results therefore suggest that coupling of the cell cycle to the circadian clock via NONO may be useful to segregate in temporal fashion cell proliferation from tissue organization.

Comparison of treatment results of acute and late injuries of the lisfranc joint.

OBJECTIVE: A retrospective comparison of treatment difficulties and treatment outcomes in Lisfranc joint injuries with late and early diagnosis. METHODS: The study group consisted of 10 patients diagnosed and treated properly within six months to 20 years of the accident causing the injury (mean six years). The control group consisted of the same number of randomly selected patients with a similar type of injury treated immediately after the accident. Mean follow-up was 13 years in the study group and eight years in the control group. The analysis evaluated the causes of the delay and the foot function at the time of follow up, measured using the AOFAS Midfoot Scale and the Lublin Foot Functional Score. The scores of the patients were analyzed using the non-parametric Mann-Whitney U test and the non-parametric Wilcoxon test. RESULTS: The control group had statistically significantly better scores on both scales. CONCLUSION: The main cause of treatment delay was misdiagnosis by the primary care physician. Level of Evidence III, Retrospective Comparative Study .

Distinct roles of DBHS family members in the circadian transcriptional feedback loop.

Factors interacting with core circadian clock components are essential to achieve transcriptional feedback necessary for metazoan clocks. Here, we show that all three members of the Drosophila behavior human splicing (DBHS) family of RNA-binding proteins play a role in the mammalian circadian oscillator, abrogating or altering clock function when overexpressed or depleted in cells. Although these proteins are members of so-called nuclear paraspeckles, depletion of paraspeckles themselves via silencing of the structural noncoding RNA (ncRNA) Neat1 did not affect overall clock function, suggesting that paraspeckles are not required for DBHS-mediated circadian effects. Instead, we show that the proteins bound to circadian promoter DNA in a fashion that required the PERIOD (PER) proteins and potently repressed E-box-mediated transcription but not cytomegalovirus (CMV) promoter-mediated transcription when they were exogenously recruited. Nevertheless, mice with one or both copies of these genes deleted show only small changes in period length or clock gene expression in vivo. Data from transient transfections show that each of these proteins can either repress or activate, depending on the context. Taken together, our data suggest that all of the DBHS family members serve overlapping or redundant roles as transcriptional cofactors at circadian clock-regulated genes.

(Re)inventing the circadian feedback loop.

For 20 years, researchers have thought that circadian clocks are defined by feedback loops of transcription and translation. The rediscovery of posttranslational circadian oscillators in diverse organisms forces us to rethink this paradigm. Meanwhile, the original "basic" feedback loops of canonical circadian clocks have swelled to include dozens of additional proteins acting in interlocked loops. We review several self-sustained clock mechanisms and propose that minimum requirements for diurnal timekeeping might be simpler than those of actual free-running circadian oscillators. Thus, complex mechanisms of circadian timekeeping might have evolved from random connections between unrelated feedback loops with independent but limited time-telling capability.

The circadian clock starts ticking at a developmentally early stage.

Although overt diurnal rhythms of behavior do not begin until well after birth, molecular studies suggest that the circadian clock may begin much earlier at a cellular level: mouse embryonic fibroblasts, for example, already possess robust clocks. By multiple criteria, we found no circadian clock present in mouse embryonic stem cells. Nevertheless, upon their differentiation into neurons, circadian gene expression was observed. In the first steps along the pathway from ES cells to neurons, a neural precursor cell (NPC) line already showed robust circadian oscillations. Therefore, at a cellular level, the circadian clock likely begins at the very earliest stages of mammalian development.

Variant alleles of the CYP1B1 gene are associated with colorectal cancer susceptibility.

BACKGROUND: CYP1B1 is a P450 enzyme which is involved in the activation of pro-carcinogens to carcinogens as well as sex hormone metabolism. Because differences in the activity of the enzyme have been correlated with variant alleles of single nucleotide polymorphisms (SNPs), it represents an attractive candidate gene for studies into colorectal cancer susceptibility. METHODS: We genotyped 597 cancer patients and 597controls for three CYP1B1 SNPs, which have previously been shown to be associated with altered enzymatic activity. Using the three SNPs, eight different haplotypes were constructed. The haplotype frequencies were estimated in cases and controls and then compared. The odds ratio for each tumour type, associated with each haplotype was estimated, with reference to the most common haplotype observed in the controls. RESULTS: The three SNPs rs10012, rs1056827 and rs1056836 alone did not provide any significant evidence of association with colorectal cancer risk. Haplotypes of rs1056827 and rs10012 or rs1056827 and rs1056836 revealed an association with colorectal cancer which was significantly stronger in the homozygous carriers. One haplotype was under represented in the colorectal cancer patient group compared to the control population suggesting a protective effect. CONCLUSION: Genetic variants within the CYP1B1 that are associated with altered function appear to influence susceptibility to a colorectal cancer in Poland. Three haplotypes were associated with altered cancer risk; one conferred protection and two were associated with an increased risk of disease. These observations should be confirmed in other populations.

[Comparison of the influence of mineral trioxide aggregate and calcium hydroxide on dental pulp of permanent teeth in biological treatment and cell cultures]. / Porównanie wplywu preparatu mineral trioxide aggregate i wodorotlenku wapnia na miazge zebóow stalych w leczeniu biologicznym i w hodowli komórkowej.

INTRODUCTION: The aim of this study was to evaluate the usefulness of a new product called Mineral Trioxide Aggregate (MTA) used in direct and indirect pulp capping. The results of pulp treatment using therapeutic MTA with the results after application of calcium hydroxide were compared. Moreover, the aim of the study was to determine the toxicity of the investigated materials in relation to human fibroblast cultures derived from dental pulp. MATERIAL AND METHODS: The study included 97 patients, male and female in the age 8-56 years. A total of 135 vital pulp therapy treatments were performed. Depending on the material and methods, the patients were assigned to four groups. RESULTS: In the carried out study 88.2% of the results were positive after a direct coverage of the pulp using MTA and 86.7% of positive results were after the application of calcium hydroxide. In the case of indirect use of both the MTA and Ca(OH)2 100% positive results were obtained. Biocompatibility test preparations showed significantly lower toxicity of MTA compared with Ca(OH)2. CONCLUSIONS: On the basis of clinical examination Mineral trioxide Aggregate proved to be as effective as calcium hydroxide in the direct and indirect pulp capping. It has been shown that the deeper damage to the pulp, of the kind during its injury, worse the prognosis compared to an exposure in the pulp. Mineral Trioxide Aggregate proved to be highly statistically significantly less toxic compared to Ca(OH)2.

BARD1 and breast cancer in Poland.

PURPOSE: To investigate whether or not a genetic variant in BARD1 (Cys557Ser) contributes to early-onset breast cancer in Poland, or modifies the risk of breast cancer in women with an inherited predisposition to breast cancer. EXPERIMENTAL DESIGN: We studied 3,188 unselected Polish women with breast cancer and 1,038 healthy controls. All women were genotyped for the BARD1 Cys557Ser variant and for known founder mutations in BRCA1 (three mutations), CHEK2 (four mutations), and NBS1 (one mutation). RESULTS: A BARD1 variant was seen in 150 of 3,188 breast cancer cases (4.7%) and in 40 of 1,038 controls (3.8%) (OR = 1.2; 95% CI = 0.9-1.7). The risk associated with the BARD1 variant was not significantly greater in women with familial cancer (OR = 1.5; 95% CI = 0.8-2.7), or with an inherited mutation in BRCA1 (OR = 0.9; 95% CI = 0.4-2.2), CHEK2 (OR = 1.0; 95% CI = 0.5-2.1), or NBS1 (OR = 1.3; 95% CI = 0.2-10.2). Modest associations were observed among the subgroups of women with very early onset breast cancer (OR = 2.9; 95% CI = 1.2-7.1) and with medullary breast cancer (OR = 1.8; 95% CI = 0.9-3.7). CONCLUSION: There was no clear association between the presence of the BARD1 Cys557Ser allele and breast cancer in Poland. Furthermore, the BARD1 Cys557Ser allele does not appear to modify the risk of breast cancers among carriers of BRCA1 mutations, or of other predisposing mutations. The allele may predispose to breast cancers of certain histologic subtypes, but further studies are needed to confirm these findings.

Circadian pattern of prolactin secretion in children with growth hormone deficiency and congenital organic lesions in the hypothalamic-pituitary region.

OBJECTIVES: Prolactin (Prl) secretion in children manifests circadian rhythm. The aim of the study was to assess circadian Prl pattern in children with growth hormone deficiency (GHD) and congenital organic disorders in the hypothalamic-pituitary region (HPR). MATERIAL AND METHODS: The analysis comprised 47 children (aged: 11.05+/-3.5 years) with GHD, divided (based on MRI) into subgroups: NORM (no disturbances in HPR); HP (pituitary hypoplasia) and PSIS (pituitary stalk interruption syndrome). The profile of circadian Prl secretion was determined, based on Prl measurements in serum every 3 hours during 24 hours. The macroscopic analysis of circadian Prl rhythm in particular groups was performed. The comparison group consists of 41 children (aged: 11.45+/-3.20 years) with idiopathic short stature (ISS). RESULTS: In GHD-HP, diurnal and nocturnal Prl concentrations were low but with the dispersion between them and with normal rhythm in most of cases. In GHD-PSIS, diurnal and nocturnal Prl concentrations were on the same level and the rhythm was not observed in most of cases. No significant differences were found in Prl secretions and Prl rhythm between GHD-NORM and ISS. The rhythm of Prl secretion was disturbed in: 72.7% of children with GHD-PSIS, 23.5% - with GHD-HP, 10.5% with GHD-NORM and 7.3% with ISS, only. CONCLUSIONS: Congenital organic lesions of HPR are associated with quantitative disorders and changes of the circadian pattern of Prl secretion. In children with GHD without organic lesions of HPR, the circadian rhythm of Prl secretion was not different from that with ISS.

Assessment of prolactin secretion in children: a profile of circadian prolactin secretion and the principles for interpreting it.

INTRODUCTION: Prolactin (Prl) is secreted in a circadian pattern, although no method of interpreting it has yet been established. The aim of the study was to assess Prl secretion in children on the basis of the Prl circadian profile and to establish principles for the interpretation of the results obtained by this method. MATERIAL AND METHODS: The analysis comprised 41 healthy short children (25 boys); aged 5.2-16.3 years, in whom hormonal disorders and chronic diseases had been excluded. The children were divided into prepubertal or pubertal subgroups. Serum Prl concentrations were measured every 3 hours for 24 hours. To assess the rhythm the parameters of macroscopic analysis were calculated and receiver operating characteristic (ROC) analysis was performed. The group for comparison consisted of 30 children aged 8.9-17.2 years with hyperprolactinaemia. RESULTS: In each subgroup significantly higher Prl concentrations were observed at night than by day. No statistical differences were noticed between the groups regarding Prl concentrations at particular time points or parameter values during circadian Prl rhythm evaluation. In the group analysed weak correlations were found between age and Prl peak and trough levels. On the basis of ROC analysis criteria for the existence of normal circadian Prl rhythm in children were established. CONCLUSIONS: 1. The presence of normal circadian Prl rhythm is observed if at least one of the following three criteria is fulfilled: amplitude >1.8779; X(n)/X(d) ratio >1.685; regression index <-0.4107. 2. No interpretation in relation to sex, age and stage of puberty is necessary for the circadian prolactin secretion rhythm in children.

CDKN2A-positive breast cancers in young women from Poland.

PURPOSE: To investigate the contribution of CDKN2A A148T common variant to early-onset breast cancer in Poland, and to establish the characteristic features of these cancers. EXPERIMENTAL DESIGN: We studied 3,069 women diagnosed with breast cancer under the age of 51 and 3,493 population controls. CDKN2A variant were detected using RFLP-PCR and confirmed by genomic sequencing. Clinical and pathological features of CDKN2A-positive cases and CDKN2A-negative cases were compared. RESULTS: A148T variant was identified in 157 of 3,069 women with breast cancer (5.1%). Overall, the odds ratio for early-onset breast cancer, given a CDKN2A variant was 1.4 (95% CI 1.075-1.725; P = 0.012). Breast tumors in women with the CDKN2A variant were more commonly intraductal cancers (DCIS) with micro-invasion (14.8% vs. 8.5%; P = 0.035). Carriers and non-carriers were similar with respect to tumor size, laterality, multicentricity, nodal status, family history and estrogen-receptor status. CONCLUSION: The CDKN2A A148T variant seems to contribute to early-onset breast cancer in Poland. Breast tumors which arise in carriers of A148T variant appear to be similar to those of breast cancers in the population at large.

CHEK2-positive breast cancers in young Polish women.

PURPOSE: To investigate the contribution of CHEK2 mutations to early-onset breast cancer in Poland and to establish the characteristic features of these cancers. EXPERIMENTAL DESIGN: We studied 3,228 women diagnosed with breast cancer under the age of 51 years and 5,496 population controls. CHEK2 mutations were detected by RFLP-PCR or allele-specific oligonucleotide-PCR assays. Clinical and pathologic features of CHEK2-positive cases and CHEK2-negative cases were compared. RESULTS: A truncating CHEK2 mutation (1100delC or IVS2+1G>A) was seen in 47 of 3,228 cases and in 34 of 5,496 controls (odds ratio, 2.4; P = 0.0001). The CHEK2 I157T missense mutation was present in 207 of 3,228 cases, compared with 264 of 5,496 controls (odds ratio, 1.4; P = 0.002). Breast cancers in women with a CHEK2 mutation were more commonly of lobular histology (21.5% versus 15.8%; P = 0.05), of size >2 cm (54.8% versus 43.5%; P = 0.01), or of multicentric origin (28.7% versus 19.5%; P = 0.01) than were cancers from women without a CHEK2 mutation. Bilateral cancers were equally common in both subgroups. CONCLUSION: Three founder alleles in CHEK2 contribute to early-onset breast cancer in Poland. Breast tumors which arise in carriers of CHEK2 mutations seem to be similar to those of breast cancers in the population at large.

Increased rates of chromosome breakage in BRCA1 carriers are normalized by oral selenium supplementation.

Women who are born with constitutional heterozygous mutations of the BRCA1 gene face greatly increased risks of breast and ovarian cancer. The product of the BRCA1 gene is involved in the repair of double-stranded DNA breaks and it is believed that increased susceptibility to DNA breakage contributes to the cancer phenotype. It is hoped therefore that preventive strategies designed to reduce chromosome damage will also reduce the rate of cancer in these women. To test for increased mutagenicity of cells from BRCA1 carriers, the frequency of chromosome breaks was measured in cultured blood lymphocytes following in vitro exposure to bleomycin in female BRCA1 carriers and was compared with noncarrier relatives. The frequency of chromosome breaks was also measured in BRCA1 carriers following oral selenium supplementation. Carriers of BRCA1 mutations showed significantly greater mean frequencies of induced chromosome breaks per cell than did healthy noncarrier relatives (0.58 versus 0.39; P < 10(-4)). The frequency of chromosome breaks was greatly reduced following 1 to 3 months of oral selenium supplementation (mean, 0.63 breaks per cell versus 0.40; P < 10(-10)). The mean level of chromosome breaks in carriers following supplementation was similar to that of the noncarrier controls (0.40 versus 0.39). Oral selenium is a good candidate for chemoprevention in women who carry a mutation in the BRCA1 gene.

[Anaplastic thyroid carcinoma in a 14-year-old boy]. / Rak anaplastyczny tarczycy u 14 letniego chlopca.

We report a case of anaplastic cancer of thyroid in 14-year-old male. The diagnosis was supported on: pace of changes, clinical image, disease course, no effect of therapy, histopathological examination.

Inhibition of cell cycle and induction of apoptosis by sulforaphane in cell lines carrying various inherited BRCA1 mutations.

The important aspect of sulforaphane (SFN) chemopreventive activity is its ability to induce cell growth inhibition and apoptosis. In this study, the effect of SFN on lymphoblastoid cells derived from people carrying four different germ-line mutations in BRCA1 gene was tested and compared to the effect of SFN on wild-type cells. The mutations studied were C61G, 3819del5, 4153delA and 5382INSC. Changes in cell viability and density after SFN treatment were evaluated, as well as cell cycle progression, changes in mitochondrial membrane potential, and phosphatidylserine externalization. SFN was shown to reduce cell viability and density in all cell lines tested. Cell cycle block in S-phase and the occurence of simultaneous apoptosis were observed. The concentration of SFN needed to elicit a comparable effect on each cell line was varied. We found that the effect of SFN on cells carrying different inherited mutations depended on mutation type.