RNA methyltransferase SPOUT1/CENP-32 links mitotic spindle organization with the neurodevelopmental disorder SpADMiSS.

SPOUT1/CENP-32 encodes a putative SPOUT RNA methyltransferase previously identified as a mitotic chromosome associated protein. SPOUT1/CENP-32 depletion leads to centrosome detachment from the spindle poles and chromosome misalignment. Aided by gene matching platforms, we identified 24 individuals with neurodevelopmental delays from 18 families with bi-allelic variants in SPOUT1/CENP-32 detected by exome/genome sequencing. Zebrafish spout1/cenp-32 mutants showed reduction in larval head size with concomitant apoptosis likely associated with altered cell cycle progression. In vivo complementation assays in zebrafish indicated that SPOUT1/CENP-32 missense variants identified in humans are pathogenic. Crystal structure analysis of SPOUT1/CENP-32 revealed that most disease-associated missense variants mapped to the catalytic domain. Additionally, SPOUT1/CENP-32 recurrent missense variants had reduced methyltransferase activity in vitro and compromised centrosome tethering to the spindle poles in human cells. Thus, SPOUT1/CENP-32 pathogenic variants cause an autosomal recessive neurodevelopmental disorder: SpADMiSS ( SPOUT1 Associated Development delay Microcephaly Seizures Short stature) underpinned by mitotic spindle organization defects and consequent chromosome segregation errors.

Molecular mechanisms of corpus callosum development: a four-step journey.

The Corpus Callosum (CC) is a bundle of axons connecting the cerebral hemispheres. It is the most recent structure to have appeared during evolution of placental mammals. Its development is controlled by a very complex interplay of many molecules. In humans it contains almost 80% of all commissural axons in the brain. The formation of the CC can be divided into four main stages, each controlled by numerous intracellular and extracellular molecular factors. First, a newborn neuron has to specify an axon, leave proliferative compartments, the Ventricular Zone (VZ) and Subventricular Zone (SVZ), migrate through the Intermediate Zone (IZ), and then settle at the Cortical Plate (CP). During the second stage, callosal axons navigate toward the midline within a compact bundle. Next stage is the midline crossing into contralateral hemisphere. The last step is targeting a defined area and synapse formation. This review provides an insight into these four phases of callosal axons development, as well as a description of the main molecular players involved.

A prospective study of the effect of testosterone escape on preradiotherapy prostate-specific antigen kinetics in prostate cancer patients undergoing neoadjuvant androgen deprivation therapy.

INTRODUCTION: Prostate-specific antigen (PSA) kinetic patterns during neoadjuvant androgen deprivation therapy have been shown to predict unfavorable long-term outcomes. OBJECTIVE: To investigate the effect of testosterone escape (TE) on these kinetic patterns, as this had not been previously reported. METHODS: There were 50 consecutive prostate cancer patients who received 6 months of triptorelin prior to definitive radiotherapy (RT). Testosterone and PSA levels were measured at baseline and every 6 weeks. Clinical factors were tested for their ability to predict for TE and unfavorable PSA kinetic patterns. The effects of TE, at both 1.7 and 0.7 nmol/L levels, were analyzed. RESULTS: TE occurred in at least one reading for 14% and 34% of the patients at the 1.7 and 0.7 nmol/L levels, respectively. No baseline factors predicted TE. The median PSA halving time was 25 days and the median pre-RT PSA level was 0.55 ng/mL. The only factor significantly associated with a higher pre-RT PSA level was a higher baseline PSA level. The only factor that significantly predicted a longer PSA halving time was TE at the 1.7 nmol/L level. CONCLUSIONS: TE and higher baseline PSA levels may adversely affect PSA kinetics and other outcomes for patients undergoing neoadjuvant hormone therapy prior to radiotherapy. Studies investigating the tailoring of neoadjuvant therapy by extending the duration in those patients with a higher baseline PSA level or by the addition of anti-androgens in those demonstrating TE, should be considered.

Is prospective MRI mapping of the changes in the volume of the prostate gland in prostate cancer patients undergoing 6 months of neo-adjuvant androgen deprivation therapy a step towards a trial to determine those who may benefit from treatment intensification or extended duration?

INTRODUCTION: Neo-adjuvant androgen deprivation therapy prior to radiotherapy (RT) causes shrinkage of the prostate gland, but the changes in volume have never been mapped over time in detail, nor have the associations between volume reduction and testosterone escape or prostate-specific antigen (PSA) kinetics been determined. METHODS: Fifty consecutive patients with prostate cancer were treated with 6 months of triptorelin prior to definitive RT. The volume of the prostate gland was measured at the outset and every 6-7 weeks thereafter using MRI scans. The volumes were calculated using a planimetric method, and inter-rater reliability was checked. Factors associated with a large initial volume and greater reductions in it were assessed. RESULTS: The median volume at the outset was 45 cc, and the median reductions every 6 weeks thereafter were 23, 18, 9 and 5%. The inter-rater agreement was high (r > 0.9, P < 0.001). There were no baseline clinical factors associated with a high initial prostate volume, but the initial volume was associated with greater volume reduction. Testosterone escape had no effect on the reduction, and changes in volume were not reflected in PSA response kinetics. CONCLUSIONS: Reductions in volume continue throughout a 6-month course of neo-adjuvant therapy but are greatest during the first 6 weeks. Although individualisation of the duration or intensity of the hormone treatment warrants further investigation, the role of prostate gland volume reduction remains uncertain. More detailed studies of tumour volume might be possible if the imaging required was acceptable and accessible to patients.

Fabrication of 1D-Nanofiber/Fe2O3 Composites with Tailored Magnetic Properties.

A novel flexible electrospun nanofiber/γ-Fe2O3 composite has been obtained from suspension of γ-Fe2O3 nanoparticles in polyvinylpyrolidone solution in dimethylformamide. The impedance spectroscopy of the synthesized nanofiber/γ-Fe2O3 composite was carried out. Negative magnetoresistance and giant magnetocapacitance effects, as well as phenomenon of a "negative capacitance" at room temperature were observed in magnetic field (2.75 κOe) in infra-low frequency range. The polarization properties and volt-ampere characteristics of the nanocomposite in the applied magnetic field indicate the increase in the dielectric permittivity ɛ and the emergence of spin electromotive force, which enables us to accumulate of electric energy at quantum level. A quantum-mechanical model, which explained the non-monotonous behaviour of the volt-ampere characteristic of the novel nanofiber based composite, has been suggested.

Comb-like PEG-containing polymeric composition as low toxic drug nanocarrier.

BACKGROUND: Development of biocompatible multifunctional polymeric drug carriers is crucial in modern pharmaceutics aimed to create "smart" drugs. The high potential of the PEGylated comb-like polymeric nanocarrier (PNC) in delivering both traditional and experimental drugs to tumor cells in vitro and in vivo has been demonstrated previously. In the present study, we investigated the general toxicity of polyethylene glycol (PEG) processed with both covalent and non-covalent attachments of PEG to compose a comb-like polymer that behaves like a simple chain of n monomers decorated with swollen side chains. The PNC possesses properties of a water-soluble surfactant containing methyl-terminated PEG side branches in some monomer units attached covalently to the carbon chain backbone. RESULTS: We have demonstrated that the synthesized PNC possesses weak toxic effects toward human leukemia cells (HL-60 and Jurkat lines), as well as toward hepatocellular (HepG2), colon (HCT116) and breast (MCF-7) tumor cell lines. Additionally, after a long period (20 days) of intraperitoneal administration, the PNC had no significant toxic effects in laboratory white mice (470 mg/kg body mass in 1 ml) and Wistar rats (440 mg/kg body mass in 10 ml). CONCLUSION: The developed PNC we studied can be qualified as a compound of grade 4 toxicity (low toxicity substance). The reduced toxicity of this PNC in combination with its improved bioavailability and previously detected capability to enhance cytotoxicity toward tumor cells in vitro and potential tumor treatment effects in vivo suggests its potential as a safe drug delivery platform for treating various diseases, especially cancer.

Which patients benefit from post-implant CT dosimetry after real-time intraoperative planning for low dose rate prostate brachytherapy? Case series and systematic literature review.

INTRODUCTION: At present, post-implant CT-based dosimetry is a standard quality assurance practice following low dose rate (LDR) prostate brachytherapy. However, it rarely influences management and involves radiation exposure, costs and inconvenience. The purpose of our study was to assess the need for post-implant CT-based dosimetry through correlation with pre-implant and real-time dosimetry and review its place in the management of patients treated with LDR brachytherapy, so that it could be undertaken more selectively. METHODS: The real-time dosimetry parameters of 34 consecutive patients who underwent LDR brachytherapy were compared with day 30 post-implant CT-based dosimetry. To validate our results against the world practice, we performed a meta-analysis of six relevant published studies, which combined data from 699 patients. The Student's t-test was performed to verify whether our dosimetric parameters significantly differ from the results of the meta-analysis. RESULTS: In our case series, the mean target volume on real-time-planned US and post-implant CT was 33.9 and 32.7 cc, respectively (P > 0.05). The dose-volume histogram (DVH) parameters were significantly different between real-time-planned and post-implant dosimetry, but re-implantation was not needed for any patients. The literature review demonstrated that there is no consensus on measures being reported. Comparison showed that our cohort had significantly smaller prostate volumes, but the DVHs were similar to other series. CONCLUSIONS: Post-implant CT and dosimetry did not alter patients' management after real-time intraoperative planning. However, we recommend that it still be employed for difficult cases or if there are any concerns identified in real-time planned dosimetry.

Hepatic metallothioneins in molecular responses to cobalt, zinc, and their nanoscale polymeric composites in frog Rana ridibunda.

Despite numerous studies suggesting a dramatic decline of amphibians, the biochemical mechanisms of adaptation in these animals to polluted environment are poorly studied. The aim of this study was to elucidate the ability to release cobalt (Co) and zinc (Zn) from their nanoscale complexes (NCs) derived from the polymeric substance of N-vinylpyrrolidone (PS) in the liver of amphibian (Rana ridibunda). Frog males were subjected to 14days exposure to waterborne Co(2+) (50µg/L), Zn(2+) (100µg/L), as well as corresponding concentrations of Co-NC, Zn-NC or PS. Main attention was paid to MT's interrelations with indices of stress and toxicity. Only Co(2+) and Zn(2+) caused elevation of the correspondent metal in MTs. Co(2+) caused down-regulation of cathepsin D activity, while Zn(2+), Zn-NC and the PS up-regulated this activity. Zn(2+) provoked 1.6 times increase of metal-bounded form of the MT (MT-Me), while all other exposures caused the elevation of the ratio of MT total protein concentration (MT-SH) and concentrations of the MT-Me and/or immunoreactive (MTi) form (up to ~10 times) accompanied by a decrease in the levels of oxyradicals. The increased DNA fragmentation and down-regulation of caspase-3 activity in relation to the redox state of glutathione and/or lactate/pyruvate were shown at all exposures. These data indicate the vulnerability of the redox state of cellular thiols and inability to release Co and Zn from NCs in frog's liver.

Responses of hepatic metallothioneins and apoptotic activity in Carassius auratus gibelio witness a release of cobalt and zinc from waterborne nanoscale composites.

The main goal of this study was to evaluate the ability of fish Carassius auratus tissues to release cobalt (Co) and zinc (Zn) cations present in the applied Co- and Zn-containing nanoscale composites (NCs). Male fish was subjected to 14day long action of Co- and Zn-NCs, as well as of Co(2+) and Zn(2+) or polymeric substance (PS) used for the NC preparation and derived from the vinylpyrrolidone. 50µg∙L(-1) of Co and 100µg∙L(-1) of Zn were applied either as a salt or a nanocomposite. Both Co and Co-NC increased (3.1 and 2.3 times, respectively) concentration of total Co, metallothionein-related Co (3.7 and 6.6 times, respectively) and thiols (by 71 and 95%, respectively), and caspase-3 activity (2.2 and 3.7 times, respectively) in the fish liver. At the same time, Co and Co-NC decreased glutathione level (1.8 and 1.9 times, respectively) and activated vitellogenesis (5.1 and 9.9 times, respectively) in the fish liver. Both Zn and Zn-NC increased markedly concentrations of metallothionein-related Zn (2.4 and 2.9 times, respectively) and Cu (2.8 and 3.2 times, respectively), and decreased metallothionein-related thiol (2.5 and 4.2 times, respectively), oxyradical (by 30.4 and 44.2%, respectively), and caspase-3 (3.0 and 5.3 times, respectively) levels in the fish liver. These peculiarities are common for metal and metal-NC and witness a release of metal from NS in fish organism. The differences in the levels of DNA strand breaks, biotransformation enzymes and total Zn levels in the liver were dependent on the kind of exposure.

Evaluation of biotargeting and ecotoxicity of Co²âº-containing nanoscale polymeric complex by applying multi-marker approach in bivalve mollusk Anodonta cygnea.

Cobalt (Co(2+)) is present in many nanoscaled materials created for various applications. The key goal of our study was to develop sensitive approaches for assessing the bio-risks associated with using novel Co(2+)-containing nanoscaled polymeric complex (Co-NC). Freshwater bivalve mollusk Anodonta cygnea (Unionidae) was subjected to 14 d action of the developed Co-NC, as well as of Co(2+) applied in the corresponding concentration (50 µg L(-1)) or polymeric substance (PS). All experimental groups under study have demonstrated signs of toxic targeting, notably changes in DNA characteristics, oxidative stress (with particularities in each exposed group) and activation of anaerobiosis (Co(2+) and Co-NC). However, the group exposed to Co-NC showed some advantages that can be related to the activation of metallothionein (MT) function (increase in the level of MT-related SH-groups (MT-SH)): low level of oxyradical formation, no increase in protein carbonylation and vitellogenin-like proteins concentration unlike in Co(2+) and PS exposed groups. On the other hand, Co(2+) increased metal (Co, Cu, Zn and Cd) binding to MT (MT-Me) without changes in MT-SH level jointly with activation of oxyradical formation and apoptosis and decreasing of lysosomal membrane stability. PS per se initiated unbalanced changes in activities of the biotransformation enzymes ethoxyresorufin-O-deethylase and glutathione-S-transferase. Thus, Co(2+) complexing with the developed PS prevented bio-toxic effects of free Co(2+) ions and PS per se, at least in the studied hydrobiont. The MT-SH was the main distinguishing index of Co-NC group selected by classification and regression tree analysis.

Smoking habits of radiotherapy patients: did the diagnosis of cancer make an impact and is there an opportunity to intervene?

OBJECTIVES: To investigate the impact of a cancer diagnosis on smoking habits in patients receiving radiotherapy and assess the opportunity to intervene. METHODS: One hundred consecutive patients were interviewed. They included patients newly diagnosed with cancer of any type and receiving radiotherapy. Detailed information was collected including smoking habits before and after the diagnosis, the timing of and reasons for any changes. We also asked about patients' view of the role of the radiation oncologist in smoking cessation and the opportunities for intervention. Analysis of results involved simple descriptive statistics. RESULTS: Although there were only 14 current smokers, only two had decided to quit. Five smokers decreased smoking, six did not change and one increased smoking. One non-smoker (1/34) took up smoking. Nearly all changes occurred within the first 30 days of diagnosis. Most (79%, 11/14) smokers believed that the treating radiation oncologist should discuss smoking cessation with their patients and that the ideal timing is at either the first consultation or when decisions about treatment have been finalised. CONCLUSIONS: The diagnosis of cancer can motivate patients to reduce smoking, but few quit altogether and a smaller number increase or even take up smoking. These changes occur early after receiving a diagnosis of cancer. Patients with a smoking history believed that the treating radiation oncologist should discuss smoking cessation with their patients and that the ideal timing is at the first consultation. The periodic nature of treatments and consultations at radiation oncology centres suggest there is the potential for an effective smoking cessation programme.