Uninfected Tissue Research Articles

Anguillicola crassus Infection Significantly Affects the Silvering Related Modifications in Steady State mRNA Levels in Gas Gland Tissue of the European Eel.

Using Illumina sequencing, transcriptional changes occurring during silvering in swimbladder tissue of the European eel have been analyzed by comparison of yellow and silver eel tissue samples. Functional annotation analysis based on GO terms revealed significant expression changes in a number of genes related to the extracellular matrix, important for the control of gas permeability of the swimbladder, and to reactive oxygen species (ROS) defense, important to cope with ROS generated under hyperbaric oxygen partial pressures. Focusing on swimbladder tissue metabolism, levels of several mRNA species encoding glucose transport proteins were several-fold higher in silver eels, while enzymes of the glycolytic pathway were not affected. The significantly higher steady state level of a transcript encoding for membrane bound carbonic anhydrase, however, suggested that CO2 production in the pentose phosphate shunt and diffusion of CO2 was of particular importance in silver eel swimbladder. In addition, the mRNA level of a large number of genes related to immune response and to sexual maturation was significantly modified in the silver eel swimbladder. The modification of several processes related to protein metabolism and transport, cell cycle, and apoptosis suggested that these changes in swimbladder metabolism and permeability were achieved by increasing cell turn-over. The impact of an infection of the swimbladder with the nematode Anguillicola crassus has been assessed by comparing these expression changes with expression changes observed between uninfected yellow eel swimbladder tissue and infected silver eel swimbladder tissue. In contrast to uninfected silver eel swimbladder tissue, in infected tissue the mRNA level of several glycolytic enzymes was significantly elevated, and with respect to extracellular matrix, several mucin genes were many-fold higher in their mRNA level. Modification of many immune related genes and of the functional categories “response to DNA damage stimulus” and “cellular response to stress” illustrated the damaging effect of the nematode infection. This study has identified a range of cellular processes in the swimbladder of silver eels that appear to be altered by nematode infection. These altered cellular processes could contribute to detrimental changes in swimbladder function that, in turn, may lead to impairment of spawning migration.

Popliteal artery aneurysms masquerading as deep vein thrombosis

A 77 year old man with known ischaemic heart disease and p paroxysmal a trial fibrillation treated with a spirin and amiodarone, presented with progressive painful swelling of his left leg. He was also known to have a 5cm abdominal aortic aneurysm, which was under surveillance. On examination, he was tachycardic and pyrexial with a tender, swollen, erythenratous left leg. A deep venous thrombosis with overlying cellulitis was diagnosed and he was commenced on intra-venous antibiotics and therapeutic doses of subcutaneous Heparin. Blood cultures grew salmonella enteriditis phage type 4 sensitive to ciprofloxacillin, which was commenced. Subsequent C T scan confirmed a large false aneurysm of the popliteal artery with surrounding soft tissue suppuration and oedema. These findings were suggestive of ruptured popliteal artery, secondary to a salmonellosis-induced mycotic aneurysm rather than deep venous thrombosis. He was taken to theatre, and a reversed long saphenous vein femoro-popliteal bypass graft was performed with the graft tunnelled subcutaneously through uninfected tissue. The intervening popliteal artery with its mycotic portion was ligated proximal and distal to the area of sepsis. Necrotic tissue in the muscle compartments of the thigh and popliteal fossa was debrided and the wound left open. Salmonella was cultured from pus specimens obtained intra-operatively. The patient made a slow recovery on the intensive care unit and was discharged back to the ward on the tenth postoperative day with a clean healing wound and a functioning bypass graft.

Determination of Ascochyta blight disease in chickpea using real-time PCR

Ascochyta blight is a devastating disease of chickpea caused by Ascochyta rabiei. In this article, we described a real-time PCR assay for the determination and quantification of A. rabiei infection in chickpea tissues and accurate monitoring of disease progression in plant materials inoculated with different inoculation methods. The primer pairs HEF1/HEF2 were designed to anneal to conserved regions of translation elongation factor 1 alpha (EF) gene for specific amplification of 82-bp fragment of A. rabiei based on SYBR Green I technology. The detection limit of assay was determined as 0.1 pg DNA. PCR specificity was confirmed by testing against uninfected chickpea tissues and another fungal species associated with chickpea. The chickpea plants were inoculated by the methods of whole-plant and detached leaflet inoculation. Disease progression in resistant and susceptible cultivars was evaluated at certain time intervals after pathogen inoculation by real-time PCR. The results revealed a good correlation between visual assessments of disease reaction and pathogen quantification in infected chickpea tissues. The target DNA sequence was also amplified from the samples of DNA extracts from artificially infested seed. This technique could provide a useful approach for efficient selection of resistant breeding material in an early stage of infection as an alternative to the visual disease assessment and will be also used for the determination and quantification of A. rabiei infection.

Erratum to: Comparative metabolomics of root nodules infected with Frankia sp. strains and uninfected roots from Alnus glutinosa and Casuarina cunninghamiana reflects physiological integration

Nitrogen-fixing root nodules on actinorhizal plants have varying internal architectures, implying diversity in how Frankia sp. integrates into plant physiologies. To understand this integration we compared the metabolomes of Alnus glutinosa and Casuarina cunninghamiana root nodules with roots from uninfected plants. High throughput gas chromatography–mass spectrometry (GC-MS) was done on extracts of nodules and roots from uninfected seedlings. Over 118 metabolites in C. cunninghamiana roots and nodules and over 163 in A. glutinosa roots and nodules were identified; between one-third to one-half of the metabolites significantly increased or decreased between roots and nodules. Amino acid patterns varied between the plants with only glutamate and alanine, which may be conducive to the induction of nitrogenase, and citrulline, elevated in nodules of both. Sugar levels were similar between species excepting a striking increase of maltose and cellobiose in C. cunninghamiana nodules indicating starch mobilization and cell wall modification. Stress related compounds increased in both systems. Phenylacetic acid was elevated in A. glutinosa nodules. High ethanolamine content was found in C. cunninghamiana nodules suggesting lipid degradation. We conclude that C. cunninghamiana responds more robustly to the presence of the endophyte than A. glutinosa with metabolite patterns consistent with different strategies used for compartmentalizing the symbiont from uninfected tissues.

Altered microRNA expression and pre-mRNA splicing events reveal new mechanisms associated with early stage Mycobacterium avium subspecies paratuberculosis infection.

The molecular regulatory mechanisms of host responses to Mycobacterium avium subsp. paratuberculosis (MAP) infection during the early subclinical stage are still not clear. In this study, surgically isolated ileal segments in newborn calves (n = 5) were used to establish in vivo MAP infection adjacent to an uninfected control intestinal compartment. RNA-Seq was used to profile the whole transcriptome (mRNAs) and the microRNAome (miRNAs) of ileal tissues collected at one-month post-infection. The most related function of the differentially expressed mRNAs between infected and uninfected tissues was “proliferation of endothelial cells”, indicating that MAP infection may lead to the over-proliferation of endothelial cells. In addition, 46.2% of detected mRNAs displayed alternative splicing events. The pre-mRNA of two genes related to macrophage maturation (monocyte to macrophage differentiation-associated) and lysosome function (adenosine deaminase) showed differential alternative splicing events, suggesting that specific changes in the pre-mRNA splicing sites may be a mechanism by which MAP escapes host immune responses. Moreover, 9 miRNAs were differentially expressed after MAP infection. The integrated analysis of microRNAome and transcriptome revealed that these miRNAs might regulate host responses to MAP infection, such as “proliferation of endothelial cells” (bta-miR-196 b), “bacteria recognition” (bta-miR-146 b), and “regulation of the inflammatory response” (bta-miR-146 b).

Sample procurement for cultures of infected wounds: a systematic review.

Identifying the responsible pathogen is required in order to select optimum antimicrobial therapy for infected wounds, but the best techniques to use remain the subject of debate. Our aim was to assess the evidence on the value of wound swabs compared with biopsies. We conducted a systematic literature review with the terms 'wound infection', 'wound swab', 'wound swab technique', 'wound biopsy', 'wound culture', 'wound swab comparison', 'Levine swab technique', 'microbiological technique', 'specimen handling', 'bacterial load', 'perioperative care', 'swab', and 'culture'. We examined yields in identifying relevant pathogens, summarised salient features of qualifying studies, and defined knowledge gaps and endpoints that future studies should address. Studies have been inconsistent, lacking specificity regarding wound types, clinical features, and sampling methods. We found moderate quality evidence that punch biopsies provide qualitative and quantitative information about the bacterial load and tissue reaction with nearly 100% sensitivity, 90% specificity and 95% accuracy for predicting wound closure. Biopsies are relatively invasive, costly, require skilled operators, and potentially exacerbate infection. Needle aspiration samples a limited portion and may enter uninfected tissue and extend infection. Wound swabs are minimally invasive, easier to perform and widely employed in clinical practice, but techniques vary. In comparative studies, the Levine technique was superior to the Z-swab techniques, and biopsies were more sensitive for antibiotic-resistant wounds than Levine or Levine-like swabs, suggesting that swabs may be useful for initial wound monitoring, but biopsies are preferred when antibiotic resistance is suspected. The Levine swab is superior to the Z-swab technique and may be useful for initial wound monitoring, but quantitative biopsies are preferred for evaluation of antibiotic-resistant wounds and to monitor the response to treatment. There is limited evidence on the role of wound swabs for detecting wound colonisation versus infection and the impact of culture-guided therapy on such clinical outcomes as eradication of infection and accelerated healing. Future studies should specify patient populations, wound types, sampling protocols, and outcomes based on culture yield and treatment results, using rigorous statistical methodology.

Albert Coons: harnessing the power of the antibody

Albert Coons: harnessing the power of the antibody

Regulatory and Functional Aspects of Indolic Metabolism in Plant Systemic Acquired Resistance

Tryptophan-derived, indolic metabolites possess diverse functions in Arabidopsis innate immunity to microbial pathogen infection. Here, we investigate the functional role and regulatory characteristics of indolic metabolism in Arabidopsis systemic acquired resistance (SAR) triggered by the bacterial pathogen Pseudomonas syringae. Indolic metabolism is broadly activated in both P. syringae-inoculated and distant, non-inoculated leaves. At inoculation sites, camalexin, indol-3-ylmethylamine (I3A), and indole-3-carboxylic acid (ICA) are the major accumulating compounds. Camalexin accumulation is positively affected by MYB122, and the cytochrome P450 genes CYP81F1 and CYP81F2. Local I3A production, by contrast, occurs via indole glucosinolate breakdown by PEN2- dependent and independent pathways. Moreover, exogenous application of the defense hormone salicylic acid stimulates I3A generation at the expense of its precursor indol-3-ylmethylglucosinolate (I3M), and the SAR regulator pipecolic acid primes plants for enhanced P. syringae-induced activation of distinct branches of indolic metabolism. In uninfected systemic tissue, the metabolic response is more specific and associated with enhanced levels of the indolics I3A, ICA, and indole-3-carbaldehyde (ICC). Systemic indole accumulation fully depends on functional CYP79B2/3, PEN2, and MYB34/51/122, and requires functional SAR signaling. Genetic analyses suggest that systemically elevated indoles are dispensable for SAR and associated systemic increases of salicylic acid. However, soil-grown but not hydroponically -cultivated cyp79b2/3 and pen2 plants, both defective in indolic secondary metabolism, exhibit pre-induced immunity, which abrogates their intrinsic ability to induce SAR.

The plant GABA signaling downregulates horizontal transfer of the Agrobacterium tumefaciens virulence plasmid.

In the tumor-inducing (Ti) Agrobacterium tumefaciens, quorum sensing activates the horizontal transfer of the virulent Ti plasmid. In pure culture, this process can be impaired by the A. tumefaciens BlcC lactonase, whose expression is induced by gamma-aminobutyrate (GABA). It was therefore hypothesized that host GABA content might modulate quorum sensing and virulence gene dissemination during A. tumefaciens infection. We examined GABA metabolism and transport in Arabidopsis thaliana tumors combining transcriptomic, metabolomic and histological approaches. In addition, using genetically modified plants and bacteria, we evaluated the impact of plant host GABA content on Ti plasmid dissemination. The results showed that GABA and free proline, which acts as an antagonist of GABA uptake in A. tumefaciens, accumulated in wild-type tumors relative to uninfected plant tissues. Moreover, comparisons of tumors induced on Col-0 and her1 plants showed that the increase in the plant GABA : proline ratio was associated with both the upregulated expression of the blcC gene and the decreased dissemination of Ti plasmid in tumor-colonizing A. tumefaciens populations. This work demonstrates experimentally that the variation in the GABA content in plant tumors can interfere with the dissemination of A. tumefaciens Ti plasmids, and therefore highlights plant GABA content as an important trait in the struggle against pathogenic bacteria.

Serotonin attenuates biotic stress and leads to lesion browning caused by a hypersensitive response to Magnaporthe oryzae penetration in rice.

The hypersensitive response (HR) of plants is one of the earliest responses to prevent pathogen invasion. A brown dot lesion on a leaf is visual evidence of the HR against the blast fungus Magnaporthe oryzae in rice, but tracking the browning process has been difficult. In this study, we induced the HR in rice cultivars harboring the blast resistance gene Pit by inoculation of an incompatible M. oryzae strain, which generated a unique resistance lesion with a brown ring (halo) around the brown fungal penetration site. Inoculation analysis using a plant harboring Pit but lacking an enzyme that catalyzes tryptamine to serotonin showed that high accumulation of the oxidized form of serotonin was the cause of the browning at the halo and penetration site. Our analysis of the halo browning process in the rice leaf revealed that abscisic acid enhanced biosynthesis of serotonin under light conditions, and serotonin changed to the oxidized form via hydrogen peroxide produced by light. The dramatic increase in serotonin, which has a high antioxidant activity, suppressed leaf damage outside the halo, blocked expansion of the browning area and attenuated inhibition of plant growth. These results suggest that serotonin helps to reduce biotic stress in the plant by acting as a scavenger of oxygen radicals to protect uninfected tissues from oxidative damage caused by the HR. The deposition of its oxide at the HR lesion is observed as lesion browning.

Protection to Fusarium oxysporum induced oxidative stress in wheat by a sub-lethal dose of Cd2+ pre-treatment.

A sub-lethal dose (50 μM) of cadmium chloride (CdCl2) pre-treatment protected wheat (Triticum aestivum, cv. Sonalika) seedlings against Fusarium oxysporum by antagonizing oxidative stress developed during infection. A reduced level of H2O2 content was observed in case of co-stressed (50 μM CdCl2 pre-treatment followed by Fusarium oxysporum infection) tissues as compared with healthy phenotypes. An elevated concentration of SA as well as DPPH, ABTS free radical scavenging activity was observed in co-stressed tissues as compared with Fusarium-infected seedlings. In contrast, lipid peroxidation and protein oxidation in co-stressed tissues were comparatively lower when compared with Fusarium-infected seedlings. However, total phenolic and chlorophyll contents were observed to be marginal in Fusarium-infected tissues as compared with co- stressed, untreated control and Cd2+ pre-treated seedlings. Thus, a comparative biochemical assay with relation to the oxidative stress in infected, uninfected and co-stressed tissues suggest the protective nature of a low dose (50 μM) of Cd2+ against Fusarium infection in wheat. Hence, a Cd2+ pre-treatment offered resistance to wheat directly and/or indirectly by reducing the oxidative stress developed during Fusarium infection.

Management of infected vascular grafts.

Infections of vascular grafts are associated with significant mortality and morbidity risk and cost an estimated $640 million annually in the United States. Clinical presentation varies by time elapsed from implantation and by surgical site. A thorough history and physical examination in conjunction with a variety of imaging modalities is often essential to diagnosis. For infected aortic grafts, there are several options for treatment, including graft excision with extra-anatomic bypass, in situ reconstruction, or reconstruction with the neo-aortoiliac system. The management of infected endovascular aortic grafts is similar. For infected peripheral bypasses, graft preservation techniques can be utilized, but in cases where it is not possible, graft removal and revascularization through uninfected tissue planes is necessary. Infected dialysis access can be surgically treated by complete or subtotal graft excision. Diagnosis, general management, and surgical approaches to infected vascular grafts are discussed in this review.

Morphological changes in experimental tuberculosis resulting from treatment with quercetin and polyvinylpyrrolidone

Research objectiveMorphological study of tissue necrosis stages in experimental organ-preserving tuberculosis pharmacotherapy using Quercetin and Polyvinylpyrrolidone (QP). Background and methods32 laboratory mice of C57BL/6JLacSto strain were used in the experiment. The animals were divided into five groups, six to seven mice in each: group 1- Mycobacterium tuberculosis (MBT) uninfected mice; group 2- MBT infected mice; group 3- MBT infected and treated with antituberculosis preparation (ATP); group 4- MBT infected and QP treated; group 5- MBT infected and treated with ATP and QP. The mice were infected through caudal vein injection with MTB H37Rv strain. The preparation QP, which belongs to the capillary-stabilizing-remedy group, was used for the research. The ATP were izoniazid and streptomycin. ResultsQP produced a strict delineation of caseous necrosis from the unaffected parts of the connective tissue with fibrosis in the center and a large number of Langerhans cells, which was not observed in the control groups without QP. The combination of QP and ATP had more pronounced effects. In MBT-infected mice, where QP was not used, unlike the group where QP was used, adipose dystrophy of hepatocytes was observed. Thus, the hepatoprotective effect of QP against TB can be suggested. ConclusionQP produces a clear delineation of caseous necrosis from an uninfected tissue by connective-tissue formation, and by forming fibrotic tissue in the center of epithelioid cells that prevents further TB dissemination by enhancing TB pharmacotherapy.

Tissue distribution of pretomanid in rat brain via mass spectrometry imaging

1. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) combines the sensitivity and selectivity of mass spectrometry with spatial analysis to provide a new dimension for histological analyses of the distribution of drugs in tissue. Pretomanid is a pro-drug belonging to a class of antibiotics known as nitroimidizoles, which have been proven to be active under hypoxic conditions and to the best of our knowledge there have been no studies investigating the distribution and localisation of this class of compounds in the brain using MALDI MSI.2. Herein, we report on the distribution of pretomanid in the healthy rat brain after intraperitoneal administration (20 mg/kg) using MALDI MSI. Our findings showed that the drug localises in specific compartments of the rat brain viz. the corpus callosum, a dense network of neurons connecting left and right cerebral hemispheres.3. This study proves that MALDI MSI technique has great potential for mapping the pretomanid distribution in uninfected tissue samples, without the need for molecular labelling.

YIA1 TBX18 Biopacemaking Improves Beating Rate and Alters Gene Expression in Bradycardic Subsidiary Right Atrial Pacemaker Tissue

Introduction Biopacemaker research aims to faithfully recapitulate robust pacemaking seen from the heart's natural pacemaker, the sinoatrial node (SAN). The transcription factor Tbx18, involved in the development of the SAN core, has previously been used to reprogramme adult ventricular myocytes into SAN-like myocytes. Subsidiary atrial pacemaker (SAP) tissue in the infero-posterior wall of the right atrium has been shown to share molecular characteristics with the SAN but is bradycardic functionally. We therefore used this SAP tissue as a model for SAN dysfunction and hypothesised that overexpressing Tbx18 in this region ectopically would increase beating rate and alter key pacemaker genes. Methods We isolated beating SAN and SAP tissues from rat right atria in vitro . We compared SAN tissue (n = 6), uninfected SAP tissue (n = 8) and SAP tissue infected with the recombinant adenovirus Ad-Tbx18 (n = 7). Beating rates were monitored during 48 h of tissue culture. We then measured mRNA levels of 14 key genes using qPCR in Ad-Tbx18 infected (n = 8) and uninfected (n = 8) SAP tissue. Data were analysed using one-way ANOVA. Results After 48 h, the mean beating rate of Ad-Tbx18 infected SAP tissue was significantly higher than uninfected SAP tissue (Ad-Tbx18: 194 ± 15.6 bpm, uninfected SAP: 142 ± 8.2 bpm; p 0.05). qPCR showed a 99.7-fold increase in Tbx18 mRNA as a result of Ad-Tbx18 infection (p < 0.001). HCN1, HCN2, RYR2, and Kv1.5 levels were significantly higher in Ad-Tbx18 infected SAP tissue than uninfected SAP tissue (p < 0.01), however HCN4 was significantly lower (p < 0.01). No significant changes were seen with Cx43, Cx45, Kir2.1, Nav1.5, NCX1, Cav1.2, Cav3.1 or Tbx3. Implications Ad-Tbx18 infection overexpressed Tbx18 in SAP tissue and increased beating rate in vitro . This led to HCN4 downregulation, but HCN1, HCN2 and RYR2 were all upregulated and this could explain the observed increase in rate. Our data raises interesting questions about a potential application of Tbx18 upregulation to the dysfunctional SAN as a biopacemaking strategy in sick sinus syndrome. ![Abstract YIA1 Figure 1][1] Abstract YIA1 Figure 1 Beating rates (mean ±SEM) were monitored between 20 and 48 h of tissue culture for isolatedsinoatrial node tissue (n = 6), uninfected inferior subsidiary pacemaker tissue(n = 8) and inferior subsidiary pacemaker tissue infected with the adenovirusAd-Tbx18 (n = 7). Rates at 48 h were significantly higher in Ad-Tbx18 infectedpreparations than uninfected preparations (p < 0.01). RT-qPCR was used toquantify mRNA for key pacemaker genes isolated after 48 h of tissue culturefrom uninfected subsidiary pacemaker tissue (n = 8) and subsidiary pacemakertissue infected with Ad-Tbx18 (n = 8). HCN1, HCN2, RYR2 and Kv1.5 were upregulatedbut HCN4 was downregulated (p < 0.05) [1]: pending:yes

T Cells Integrate Local and Global Cues to Discriminate between Structurally Similar Antigens.

T lymphocytes’ ability to discriminate between structurally related antigens has been attributed to the unique signaling properties of the T cell receptor. However, recent studies have suggested that the output of this discrimination process is conditioned by environmental cues. Here, we demonstrate how the IL-2 cytokine, collectively generated by strongly activated T cell clones, can induce weaker T cell clones to proliferate. We identify the PI3K pathway as being critical for integrating the antigen and cytokine responses and for controlling cell-cycle entry. We build a hybrid stochastic/deterministic computational model that accounts for such signal synergism and demonstrates quantitatively how T cells tune their cell-cycle entry according to environmental cytokine cues. Our findings indicate that antigen discrimination by T cells is not solely an intrinsic cellular property but rather a product of integration of multiple cues, including local cues such as antigen quality and quantity, to global ones like the extracellular concentration of inflammatory cytokines.

Macrophages are required for host survival in experimental urogenital schistosomiasis.

Urogenital schistosomiasis, Schistosoma haematobium worm infection, afflicts millions of people with egg-triggered, fibrotic bladder granulomata. Despite the significant global impact of urogenital schistosomiasis, the mechanisms of bladder granulomogenesis and fibrosis are ill defined due to the prior lack of tractable animal models. We combined a mouse model of urogenital schistosomiasis with macrophage-depleting liposomal clodronate (LC) to define how macrophages mediate bladder granulomogenesis and fibrosis. Mice were injected with eggs purified from infected hamsters or vehicle prepared from uninfected hamster tissues (xenoantigen and injection trauma control). Empty liposomes were controls for LC: 1) LC treatment resulted in fewer bladder egg granuloma-infiltrating macrophages, eosinophils, and T and B cells, lower bladder and serum levels of eotaxin, and higher bladder concentrations of IL-1α and chemokines (in a time-dependent fashion), confirming that macrophages orchestrate leukocyte infiltration of the egg-exposed bladder; 2) macrophage-depleted mice exhibited greater weight loss and bladder hemorrhage postegg injection; 3) early LC treatment postegg injection resulted in profound decreases in bladder fibrosis, suggesting differing roles for macrophages in fibrosis over time; and 4) LC treatment also led to egg dose-dependent mortality, indicating that macrophages prevent death from urogenital schistosomiasis. Thus, macrophages are a potential therapeutic target for preventing or treating the bladder sequelae of urogenital schistosomiasis.

Fate map of Medicago truncatula root nodules.

Legume root nodules are induced by N-fixing rhizobium bacteria that are hosted in an intracellular manner. These nodules are formed by reprogramming differentiated root cells. The model legume Medicago truncatula forms indeterminate nodules with a meristem at their apex. This organ grows by the activity of the meristem that adds cells to the different nodule tissues. In Medicago sativa it has been shown that the nodule meristem is derived from the root middle cortex. During nodule initiation, inner cortical cells and pericycle cells are also mitotically activated. However, whether and how these cells contribute to the mature nodule has not been studied. Here, we produce a nodule fate map that precisely describes the origin of the different nodule tissues based on sequential longitudinal sections and on the use of marker genes that allow the distinction of cells originating from different root tissues. We show that nodule meristem originates from the third cortical layer, while several cell layers of the base of the nodule are directly formed from cells of the inner cortical layers, root endodermis and pericycle. The latter two differentiate into the uninfected tissues that are located at the base of the mature nodule, whereas the cells derived from the inner cortical cell layers form about eight cell layers of infected cells. This nodule fate map has then been used to re-analyse several mutant nodule phenotypes. This showed, among other things, that intracellular release of rhizobia in primordium cells and meristem daughter cells are regulated in a different manner.

Level of Estrogens and Their Metabolites in Infected Ovarian Tissue in Women Having Uterine Carcinomas

ABSTRACT Aim: Recently growing evidence has indicated the presence of persistent viruses and microorganisms in malignant tumor cells. Studying of the effect of infectious factors on the changes in endocrine profile in hormone-dependent tissues can contribute to detection of the reasons of sharp rise in the frequency of cancer in women over fifty years and the possibility of prevention and treatment of oncopathologies. Methods: 67 samples of ovarian tissue in patients with uterine carcinomas T1-2N0M0 were studied, mean age 53,4±3,2 years. Levels of estrone (Е1), estradiol (Е2), estriol (Е3), estrogen metabolite ratio 2-ОНЕ and 16-ОНЕ were detected in 10% homogenates by enzyme-linked immunosorbent assay with the help of standard test systems (XEMA Co., Ltd.). Intact ovarian tissue without IgG antibodies against infectious agents served as a control. Polymerase chain reaction method was used for the detection of cytomegalovirus (CMV), herpes simplex virus (HSV) and chlamydia infection in ovarian tissue. Statistical analysis of results was made using Statistica 6.0 programme. Written voluntary informed consent of all patients was received in all cases. Results: 35,8% of 67 samples of ovarian tissue were infected with complex of infections – clamydias+HSV+CMV; 32,8% – HSV only; 11,9% – CMV+HSV; 11,9% – CMV only. Only 7,6% of the samples occurred to be intact. The results of the study show increase of Е1 concentration by 1,6-3,5 times and decrease of Е2 by 7,3-16,4 times in infected ovaries in comparison with uninfected tissues. When calculating the ratio Е1/Е2, maximum prevalence of Е1 – by 25-28,8 times higher in comparison with uninfected tissue – was recorded in HSV and HSV+CMV. In all other cases – CMV and clamydias+CMV – Е1/Е2 ratio increased by 15-16 times. Estrogen metabolite ratio 2-ОНЕ/16-ОНЕ in infected ovarian tissue was decreased on average by 1,5 times in comparison with intact samples. Conclusions: Е1/Е2 ratio changes towards Е1 prevalence in infected ovarian tissue, and estrogen metabolite ratio decreases towards aggressive 16ОНЕ prevalence. Disclosure: All authors have declared no conflicts of interest.

Inhibition of placental mTORC1 signalling may explain decreased placental nutrient transport and contribute to restricted fetal growth in placental malaria

Inhibition of placental mTORC1 signalling may explain decreased placental nutrient transport and contribute to restricted fetal growth in placental malaria