Immunohistochemical analysis of NaPi2b protein (MX35 antigen) expression and subcellular localization in human normal and cancer tissues

Aim: To study the expression profile of the NaPi2b protein and its localization in breast, ovarian and lung cancer cells in relation to normal tissues adjacent to tumor. Methods: Immunohistochemical analysis with monoclonal antibody MX35 was applied for investigation of NaPi2b protein expression in breast, lung and ovarian carcinomas. Intensity of NaPi2b protein expression was calculated with semiquantitative scores. Results: NaPi2b (MX35) protein expression was detected in breast, lung and ovarian cancer cells and adjacent normal tissue. We have shown that in contrast to ovarian tumors in breast and lung tumors NaPi2b expression is down regulated comparing to correspondent normal tissues. Conclusion: This study provides the data on the pattern of NaPi2b expression and cellular localization in breast, lung and ovarian cancers, which might be useful for understanding the mechanism of transport and maintenance of inorganic phosphate in cancer and normal cells, as well as for developing novel immunotherapeutic approaches based on MX35 monoclonal antibody

Digital photographic technique for the production of an artificial eye

The use of hand painting an iris button using oil paint remains the conventional method of artificial eye manufacturing. The authors found that replacing this technique with a digital photograph taken of a patient’s unaffected eye offers several advantages over the conventional method but the process from capture to print must be standardised and colour accurate. The authors of this paper suggest a tried and tested formulated photographic process of capture and printing prior to polymerisation. It discusses issues that can arise and how these can be overcome in order to achieve a high-quality print that can be used to produce a ‘life like’ ocular prosthesis

Difference between vinblastine and vincristine in distribution in the blood of rats and binding by platelets and malignant cells

Abstract--Tritium labeled vinblastine ( VLB ) and vincristine ( VC

Using small molecules to facilitate exchange of bicarbonate and chloride anions across liposomal membranes

Bicarbonate is involved in a wide range of biological processes, which include respiration, regulation of intracellular pH and fertilization. In this study we use a combination of NMR spectroscopy and ion-selective electrode techniques to show that the natural product prodigiosin, a tripyrrolic molecule produced by microorganisms such as Streptomyces and Serratia, facilitates chloride/bicarbonate exchange (antiport) across liposomal membranes. Higher concentrations of simple synthetic molecules based on a 4,6-dihydroxyisophthalamide core are also shown to facilitate this antiport process. Although it is well known that proteins regulate Cl-/HCO3- exchange in cells, these results suggest that small molecules may also be able to regulate the concentration of these anions in biological systems

Novel artificial eye service evaluation using patient reported outcome measures

Background This service evaluation explores patient reported outcomes from patients provided with high definition ocular prostheses (artificial eyes). Methods Validated patient questionnaires (FACE-Q, DAS24 and HADS) were utilised to evaluate patient experiences of their new ocular prosthesis. 10 patients were included in the service evaluation, which was conducted between December 2018 and September 2019. Descriptive analysis of the mean and 95% CI was undertaken for all questionnaires. Statistical analysis was performed using SPSS 21 Principal Component Analysis (PCA) for FACE-Q questionnaires. Correlations were significant when factor loading is at α > 0.4. Results A questionnaire response rate of 80% was achieved (n = 8). PCA analysis showed the number of variables tested could be reduced. Two principal components (PC1 and PC2) had very good to excellent internal consistency between variables with factor loading (α = 0.7–0.9). PC1 contained questionnaires 1–7, all of which were highly correlated. PC2 contained question number 8 with a factor loading of α = 0.8. This indicates good reliability, validity and responsiveness. Conclusions We hope to demonstrate the importance of service evaluations with respect to rapidly evolving technological advances in medical devices, pharmaceuticals and imaging modalities. Further feasibility and full clinical studies are required to confirm the positive results of the novel artificial eye service we have evaluated with respect to the traditional approach

Диференціація генотипів сосни звичайної за поліморфізмом довжини інтронів генів дефензинів

The search for highly informative DNA markers to support the breeding programs aimed at increasing the productivity and biological stability of forest stands is an urgent task, especially in the context of global climate change. In this paper, we first investigate a new type of genetic markers based on the intron length polymorphism (ILP) of defensin genes to determine their informativeness and promising use for the estimation of the potential resistance of pine genotypes to biological damage. In the course of our study we applied the following methods: total DNA isolation by STAB method, PCR amplification with specific primers to pine defensin genes and Heterobasidion annosum s. s., electrophoretic separation of PCR fragments on a polyacrylamide gel under nondenaturing conditions, and also data analysis using software GenAlEx 6.053 and Statistica 10. Having conducted the research, we can present the following results. To investigate the defensin genes polymorphism in pine trees, we used ILP markers. These markers were developed based on the nucleotide sequences of the exons of PsDef1-4 genes. Analysis of PCR fragments obtained after amplification of each pair of primers with genomic DNA of pine showed that only one pair of primers, which is specific to defensin 2 (IPL-PsDef2), forms a wide range of amplified products, indicating their promising use for genetic characterization of genotypes of Scots pine. To clarify the use of IPL-PsDef2 markers for the study of genetic polymorphism we analyzed 196 trees and revealed that the average PIC was 0.287. IPL-PsDef2 markers were used to analyze the different genotypes of Scots pine on the areas affected by root rot disease. Based on the results of cluster analysis, the samples were divided into two groups, which differ in resistance to the root sponge. In addition, we identified PCR fragments that are specific to each of these groups and can serve as markers for the evaluation of the resistance of pine genotypes to Heterobasidion annosum. Thus, our conclusion is as follows: genotyping of defensin genes loci of pine breeding material is promising for the development of environmentally friendly technologies in order to enhance the sustainability of improved genetic material of pine trees to biotic stress.Наведено результати дослідження генотипів сосни звичайної (Pinus sylvestris L.) із використанням молекулярних маркерів на підстаі генетичного поліморфізму довжини інтронів генів дефензинів PsDef1-4. Виявлено високу інформативність маркерів на підстаі інтрону гена PsDef2 (IPL-PsDef2) для дослідження внутрішньовидового поліморфізму в сосни звичайної. Встановлено межі осередка інфекції кореневої губки (Heterobasidion annosum (Fr.) Bref.) в сосновому насадженні за допомогою полімеразно-ланцюгової реакції (ПЛР) із використанням видоспецифічних праймерів. Проведено ДНК-профілювання модельних дерев способом електрофоретичного розділення ампліконів IPL-PsDef2 маркерів. За результатами поліморфізму довжини інтронів генів дефензинів здійснено кластеризацію генотипів сосни звичайної із використанням алгоритму UPGMA. на підстаі дендрограми виділено два кластери генотипів зі 100 % бутстреп підтримкою, які відрізняються за стійкістю до кореневої губки. Методом К-середніх визначено значущість кожного з ампліконів для диференціації генотипів сосни на групи. Виявлено амплікони, які можуть бути генетичними маркерами для оцінки потенційної стійкості генотипів сосни звичайної до кореневої губки. Отримані дані щодо поліморфізму локусів генів дефензинів, а також результати кластерного аналізу вказують на перспективність використання IPL-PsDef2 маркерів для генотипування сосни звичайної

The xc− cystine/glutamate antiporter: a mediator of pancreatic cancer growth with a role in drug resistance

The xc− cystine transporter enhances biosynthesis of glutathione, a tripeptide thiol important in drug resistance and cellular defense against oxidative stress, by enabling cellular uptake of cystine, a rate-limiting precursor. Because it is known to regulate glutathione levels and growth of various cancer cell types, and is expressed in the pancreas, we postulate that it is involved in growth and drug resistance of pancreatic cancer. To examine this, we characterised expression of the xc− transporter in pancreatic cancer cell lines, MIA PaCa-2, PANC-1 and BxPC-3, as subjected to cystine-depletion and oxidative stress. The results indicate that these cell lines depend on xc−-mediated cystine uptake for growth, as well as survival in oxidative stress conditions, and can modulate xc− expression to accommodate growth needs. Immunohistochemical analysis showed that the transporter was differentially expressed in normal pancreatic tissues and overexpressed in pancreatic cancer tissues from two patients. Furthermore, gemcitabine resistance of cells was associated with elevated xc− expression and specific xc− inhibition by monosodium glutamate led to growth arrest. The results suggest that the xc− transporter by enhancing glutathione biosynthesis plays a major role in pancreatic cancer growth, therapy resistance and represents a potential therapeutic target for the disease

Extensive Anti-CoA Immunostaining in Alzheimer’s Disease and Covalent Modification of Tau by a Key Cellular Metabolite Coenzyme A

Alzheimer’s disease (AD) is a neurodegenerative disorder, accounting for at least two-thirds of dementia cases. A combination of genetic, epigenetic and environmental triggers is widely accepted to be responsible for the onset and development of AD. Accumulating evidence shows that oxidative stress and dysregulation of energy metabolism play an important role in AD pathogenesis, leading to neuronal dysfunction and death. Redox-induced protein modifications have been reported in the brain of AD patients, indicating excessive oxidative damage. Coenzyme A (CoA) is essential for diverse metabolic pathways, regulation of gene expression and biosynthesis of neurotransmitters. Dysregulation of CoA biosynthesis in animal models and inborn mutations in human genes involved in the CoA biosynthetic pathway have been associated with neurodegeneration. Recent studies have uncovered the antioxidant function of CoA, involving covalent protein modification by this cofactor (CoAlation) in cellular response to oxidative or metabolic stress. Protein CoAlation has been shown to both modulate the activity of modified proteins and protect cysteine residues from irreversible overoxidation. In this study, immunohistochemistry analysis with highly specific anti-CoA monoclonal antibody was used to reveal protein CoAlation across numerous neurodegenerative diseases, which appeared particularly frequent in AD. Furthermore, protein CoAlation consistently co-localized with tau-positive neurofibrillary tangles, underpinning one of the key pathological hallmarks of AD. Double immunihistochemical staining with tau and CoA antibodies in AD brain tissue revealed co-localization of the two immunoreactive signals. Further, recombinant 2N3R and 2N4R tau isoforms were found to be CoAlated in vitro and the site of CoAlation mapped by mass spectrometry to conserved cysteine 322, located in the microtubule binding region. We also report the reversible H_{2}O_{2}-induced dimerization of recombinant 2N3R, which is inhibited by CoAlation. Moreover, CoAlation of transiently expressed 2N4R tau was observed in diamide-treated HEK293/Pank1β cells. Taken together, this study demonstrates for the first time extensive anti-CoA immunoreactivity in AD brain samples, which occurs in structures resembling neurofibrillary tangles and neuropil threads. Covalent modification of recombinant tau at cysteine 322 suggests that CoAlation may play an important role in protecting redox-sensitive tau cysteine from irreversible overoxidation and may modulate its acetyltransferase activity and functional interactions

Allelic Lineages of the Ficolin Genes (FCNs) Are Passed from Ancestral to Descendant Primates

The ficolins recognize carbohydrates and acetylated compounds on microorganisms and dying host cells and are able to activate the lectin pathway of the complement system. In humans, three ficolin genes have been identified: FCN1, FCN2 and FCN3, which encode ficolin-1, ficolin-2 and ficolin-3, respectively. Rodents have only two ficolins designated ficolin-A and ficolin-B that are closely related to human ficolin-1, while the rodent FCN3 orthologue is a pseudogene. Ficolin-2 and ficolin-3 have so far only been observed in humans. Thus, we performed a systematic investigation of the FCN genes in non-human primates. The exons and intron-exon boundaries of the FCN1-3 genes were sequenced in the following primate species: chimpanzee, gorilla, orangutan, rhesus macaque, cynomolgus macaque, baboon and common marmoset. We found that the exon organisation of the FCN genes was very similar between all the non-human primates and the human FCN genes. Several variations in the FCN genes were found in more than one primate specie suggesting that they were carried from one species to another including humans. The amino acid diversity of the ficolins among human and non-human primate species was estimated by calculating the Shannon entropy revealing that all three proteins are generally highly conserved. Ficolin-1 and ficolin-2 showed the highest diversity, whereas ficolin-3 was more conserved. Ficolin-2 and ficolin-3 were present in non-human primate sera with the same characteristic oligomeric structures as seen in human serum. Taken together all the FCN genes show the same characteristics in lower and higher primates. The existence of trans-species polymorphisms suggests that different FCN allelic lineages may be passed from ancestral to descendant species

Low Amplitude Boom-and-Bust Cycles Define the Septoria Nodorum Blotch Interaction

Introduction: Septoria nodorum blotch (SNB) is a complex fungal disease of wheat caused by the Dothideomycete fungal pathogen Parastagonospora nodorum. The fungus infects through the use of necrotrophic effectors (NEs) that cause necrosis on hosts carrying matching dominant susceptibility genes. The Western Australia (WA) wheatbelt is a SNB “hot spot” and experiences significant under favorable conditions. Consequently, SNB has been a major target for breeders in WA for many years. Materials and Methods: In this study, we assembled a panel of 155 WA P. nodorum isolates collected over a 44-year period and compared them to 23 isolates from France and the USA using 28 SSR loci. Results: The WA P. nodorum population was clustered into five groups with contrasting properties. 80% of the studied isolates were assigned to two core groups found throughout the collection location and time. The other three non-core groups that encompassed transient and emergent populations were found in restricted locations and time. Changes in group genotypes occurred during periods that coincided with the mass adoption of a single or a small group of widely planted wheat cultivars. When introduced, these cultivars had high scores for SNB resistance. However, the field resistance of these new cultivars often declined over subsequent seasons prompting their replacement with new, more resistant varieties. Pathogenicity assays showed that newly emerged isolates non-core are more pathogenic than old isolates. It is likely that the non-core groups were repeatedly selected for increased virulence on the contemporary popular cultivars. Discussion: The low level of genetic diversity within the non-core groups, difference in virulence, low abundance, and restriction to limited locations suggest that these populations more vulnerable to a population crash when the cultivar was replaced by one that was genetically different and more resistant. We characterize the observed pattern as a low-amplitude boom-and-bust cycle in contrast with the classical high amplitude boom-and-bust cycles seen for biotrophic pathogens where the contrast between resistance and susceptibility is typically much greater. Implications of the results are discussed relating to breeding strategies for more sustainable SNB resistance and more generally for pathogens with NEs