Cannabinoid receptors expression in bone marrow trephine biopsy of chronic lymphocytic leukaemia patients treated with purine analogues

Background: Cannabinoid receptors CB1 and CB2 are part the endocannabinoid system that plays an important role in the process of proliferation and apoptosis of different neoplastic cells. B-cell chronic lymphocytic leukaemia is one of the diseases in which these processes are altered. Aim: The aim of our study was the assessment of cannabinoid receptor expression on the B-lymphocytes in bone marrow trephine biopsy from leukaemic patients at diagnosis and after purine analogue treatment. Methods: The biopsy was taken routinely and standard immunohistochemical staining procedure for paraffin embedded sections was applied. The cannabinoid receptors were detected using specific primary polyclonal antibody anti-CB1 and anti-CB2. Additionally, an existence of cannabinoid receptors was confirmed by flow cytometry. Results: The results showed that the expression of CB1 receptor on the surface of neoplastic cells was lower than that of CB2 (17.0 ± 3.1% and 92.1 ± 1.7% respectively, p < 0.001). Nine of the patients responded to applied treatment with a reduction in leukaemic infiltration (77.2 ± 6.9% to 30.2 ± 6.5%, p = 0.007) and CB1 receptor expression (24.4 ± 4.8% to 8.6 ± 2.9%, p = 0.01), but there was no change in CB2 expression (91.7 ± 2.7% vs 90.9 ± 2.8%, p = 0.69). Four patients without remission expressed even greater number of the receptors. In all of the cases both cannabinoid receptor types antibodies gave positive reaction. Furthermore, the existence of cannabinoid receptors on neoplastic lymphocytes was confirmed by flow cytometry. Conclusion: The study provides original evidence for the existence of cannabinoid receptors on B-lymphocytes in chronic lymphocytic leukaemia patients. The receptors are thought to be a new structure that can modify the course of the disease and may be considered as a new target in leukaemia treatment.Обоснование: рецепторы каннабиноидов CB1 и CB2 яв ляются частью системы эндоканнабиноидов, которая играет важную роль в процессах пролиферации и апоптоза различных неопластических клеток. Одним из заболеваний, при которых происходит нарушение этих процессов, является В-клеточный хронический лимфлейкоз. Цель: оценка экспрессии рецепторов каннабиноидов на В-лимфоцитах в трепанобиоптатах у больных лейкозом до и после проведения лечения с использованием пуриновых аналогов. Методы: биоптаты получили рутинными методами; иммуногистохимическое исследование депарафинизированных срезов проводили по стандартной процедуре. Рецепторы каннабиноидов определяли с использованием специфических моноклональных антител анти-CB1 и анти-CB2. Кроме того, наличие рецепторов подтверждено при по- мощи проточной цитофлуориметрии. Результаты: показано, что экспрессия рецептора CB1 на поверхности опухолевых клеток ниже, чем экспрессия CB2 (17,0  ± 3,1% и 92,1 1,7% твенно, p < 0,001). После проведенного лечения у 9 пациентов отмечалось уменьшение лейкозного инфильтрата (77,2 6,9% до 30,2 6,5%, p = 0,007) и снижение экспрессии рецептора CB1 (24,4 4,8% до 8,6 2,9%, p = 0,01), однако различий в экспрессии CB2 не отмечали (91,7 2,7% против 90,9 2,8%, p = 0,69). У 4 пациентов, у которых не удалось достичь ремиссии, определяли даже повышение экспрессии рецепторов. Во всех случаях маркировки антителами к обоим типам рецепторов каннабиноидов отмечали поло- жительную реакцию. Более того, присутствие рецепторов каннабиноидов на злокачественных клетках подтверждали при помощи проточной цитометрии. Выводы: в ходе исследования показано изначальное наличие рецепторов каннабиноидов на В-лимфоцитах у больных хроническим лимфолейкозом. Указанные рецепторы могут быть новой структурой, которая может быть модифицирована в течение болезни, и могут считаться новой мишенью при лечении больных лейкозом

Transcriptome Sequencing Reveals Novel Candidate Genes for Cardinium hertigii-Caused Cytoplasmic Incompatibility and Host-Cell Interaction

Cytoplasmic incompatibility (CI) is an intriguing, widespread, symbiont-induced reproductive failure that decreases offspring production of arthropods through crossing incompatibility of infected males with uninfected females or with females infected with a distinct symbiont genotype. For years, the molecular mechanism of CI remained unknown. Recent genomic, proteomic, biochemical, and cell biological studies have contributed to understanding of CI in the alphaproteobacterium Wolbachia and implicate genes associated with the WO prophage. Besides a recently discovered additional lineage of alphaproteobacterial symbionts only moderately related to Wolbachia, Cardinium (Bacteroidetes) is the only other symbiont known to cause CI, and genomic evidence suggests that it has very little homology with Wolbachia and evolved this phenotype independently. Here, we present the first transcriptomic study of the CI Cardinium strain cEper1, in its natural host, Encarsia suzannae, to detect important CI candidates and genes involved in the insect-Cardinium symbiosis. Highly expressed transcripts included genes involved in manipulating ubiquitination, apoptosis, and host DNA. Female-biased genes encoding ribosomal proteins suggest an increase in general translational activity of Cardinium in female wasps. The results confirm previous genomic analyses that indicated that Wolbachia and Cardinium utilize different genes to induce CI, and transcriptome patterns further highlight expression of some common pathways that these bacteria use to interact with the host and potentially cause this enigmatic and fundamental manipulation of host reproduction

Hippocampal and diencephalic pathology in developmental amnesia.

Developmental amnesia (DA) is a selective episodic memory disorder associated with hypoxia-induced bilateral hippocampal atrophy of early onset. Despite the systemic impact of hypoxia-ischaemia, the resulting brain damage was previously reported to be largely limited to the hippocampus. However, the thalamus and the mammillary bodies are parts of the hippocampal-diencephalic network and are therefore also at risk of injury following hypoxic-ischaemic events. Here, we report a neuroimaging investigation of diencephalic damage in a group of 18 patients with DA (age range 11-35 years), and an equal number of controls. Importantly, we uncovered a marked degree of atrophy in the mammillary bodies in two thirds of our patients. In addition, as a group, patients had mildly reduced thalamic volumes. The size of the anterior-mid thalamic (AMT) segment was correlated with patients' visual memory performance. Thus, in addition to the hippocampus, the diencephalic structures also appear to play a role in the patients' memory deficit

Electrostatically gated membrane permeability in inorganic protocells

Although several strategies are now available to produce functional microcompartments analogous to primitive cell-like structures, little progress has been made in generating protocell constructs with self-controlled membrane permeability. Here we describe the preparation of water-dispersible colloidosomes based on silica nanoparticles and delineated by a continuous semipermeable inorganic membrane capable of self-activated, electrostatically gated permeability. We use crosslinking and covalent grafting of a pH-responsive copolymer to generate an ultrathin elastic membrane that exhibits selective release and uptake of small molecules. This behaviour, which depends on the charge of the copolymer coronal layer, serves to trigger enzymatic dephosphorylation reactions specifically within the protocell aqueous interior. This system represents a step towards the design and construction of alternative types of artificial chemical cells and protocell models based on spontaneous processes of inorganic self-organization

Selection for Replicases in Protocells

PMCID: PMC3649988This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Hippocampal damage and memory impairment in congenital cyanotic heart disease

Neonatal hypoxia can lead to hippocampal atrophy, which can lead, in turn, to memory impairment. To test the generalizability of this causal sequence, we examined a cohort of 41 children aged 8-16, who, having received the arterial switch operation to correct for transposition of the great arteries, had sustained significant neonatal cyanosis but were otherwise neurodevelopmentally normal. As predicted, the cohort had significant bilateral reduction of hippocampal volumes relative to the volumes of 64 normal controls. They also had significant, yet selective, impairment of episodic memory as measured by standard tests of memory, despite relatively normal levels of intelligence, academic attainment, and verbal fluency. Across the cohort, degree of memory impairment was correlated with degree of hippocampal atrophy suggesting that even as early as neonatal life no other structure can fully compensate for hippocampal injury and its special role in serving episodic long term memory

Methyl paraben as a sex pheromone in canine urine – is the question still open?

The literature concerning the issue of canine sex pheromones includes reports presenting completely conflicting opinions about the chemical composition of the canine urine in the context of semiochemical communication. At present, the predominant report cited by many different authors is the article published in Science in 1979 by Goodwin at al., presenting methyl p-hydroxybenzoate (methyl paraben) as the main canine sex pheromone. While it has been proved that pure methyl paraben lacks semiochemical activity as do commercially available products containing this substance (Eau D’Estrus, Synbiotics, USA), in view of the conflicting published reports the aim of this study was to revaluate using modern techniques the presence of methyl p-hydroxybenzoate in canine urine during different phases of the ovarian cycle. Ten female dogs of different breeds were used. Urine samples from bitches collected during various stages of the ovarian cycle were examined with using the SPME and GC/MS methods. Methyl paraben was not detected in any of the samples. In conclusion, because of the lack of methyl-p-hydroxybenzoate in the samples examined, the present study confirmed negative opinions on the possibility of this substance playing a crucial role in semiochemical communication during reproduction in dogs (Canis familiaris)