Bulb Tissue Research Articles

Accumulation of heavy metal in onion (Allium cepa) plants irrigated with treated wastewater under field conditions

Accumulation of heavy metals in edible parts of crops irrigated with treated wastewater could create a health hazard. The objective of the study was to assess the distribution of heavy metals in edible and non-edible parts of onion (Allium cepa) plants irrigated with treated wastewater under field conditions. A 15-ha field was established for onions irrigated with centre pivot using commercial cultural practices. At harvest, 1 ha among the 15 ha was selected at random and demarcated into eight blocks for sampling five plants at random during two different seasons. Plants were separated into root, bulb and leaf, which were separately dried, ground and quantified for heavy metals using ICP-OES. In Experiment 1, plant organ treatments were significant (P<0.05) on Arsenic (As), Manganese (Mn), Cadmium (Cd), Cobalt (Co), Chromium (Cr), Copper (Cu) and Lead (Pb), contributing 73, 84, 80, 76, 95 and 81% in total treatment variation (TTV), of the respective treatments. In Experiment 2, treatments were significant on As, Mn, Cd, Cr, Cu and Pb, contributing 83, 88, 80, 74, 96 and 78% in TTV of the respective variables, with no significant effects on Co. Relative to root tissues, the accumulation of heavy metals increased (32–35%) and decreased (20–87%) in leaf and bulb tissues, respectively. In conclusion, onion bulbs were relatively safe for consumption in plants irrigated with treated wastewater.

Особливості кореляції рівня функції та спастичності паретичної кінцівки за різних видів тканинної нейротрансплантації на моделі спінальної травми

Мета роботи: дослідити кореляцію рівня спастичності та функції паретичної кінцівки за різних видів тканинної нейротрансплантації на моделі спінальної травми.Матеріали і методи. Тварини – білі безпородні щури-самці (5,5 міс, ~350 г), групи: “контроль” – лівобічний перетин половини поперечника спинного мозку на рівні Т11 (n=16), “ТТНЦ” – аналогічна травма + негайна трансплантація тканини нюхової цибулини зрілого щура в зону ураження спинного мозку (n=34); “ТТФМ” – трансплантація тканини фетального мозочка щура (Е18) за аналогічних умов (n=15); “ТТФН” — трансплантація тканини фетальної нирки щура (Е18) за аналогічних умов (n=8). Дослідження показника функції (ПФ) та показника спастичності (ПС) задньої іпсилатеральної кінцівки (ЗІК) – шкала Basso–Beattie–Bresnahan (ВВВ) та шкала Ashworth відповідно.Результати дослідження та їх обговорення. ТТНЦ забезпечує достовірну перевагу ПФ ЗІК із максимумом на 3-му тижні (3,7±0,5 бала ВВВ) з подальшим поступовим зменшенням до (2,4±0,6) бала ВВВ (24-й тиждень), зменшує вираження спастичності у ранньому періоді травми. ТТФМ перетворює динаміку ПФ ЗІК з прогредієнтної у константну з коливаннями ПФ ЗІК у межах 3–3,6 бала ВВВ протягом усього експерименту, достовірно підвищує рівень спастичності протягом 1–3-го тижня, у подальшому стабілізує ПС ЗІК на рівні 1,8–2,1 бала Ashworth. ТТФН зумовлює наявність прогредієнтної (1-й місяць) та прогредієнтно-константної (6–24-й тиждень) фаз динаміки ПФ ЗІК, достовірно потенціюючи спастичність протягом перших двох тижнів процесу. У динаміці спостереження характерна відсутність кореляції між середніми значеннями ПФ та ПС ЗІК для групи “ТТФМ”, слабка додатна кореляція для групи “контроль”, помірна додатна кореляція для групи “ТТФН”, сильна від’ємна кореляція для групи “ТТНЦ”. Кореляційний аналіз індивідуальних значень ПФ та ПС ЗІК впродовж експерименту та на кожному з термінів спостереження свідчить про переважання від’ємної кореляції між ними.Висновок. Варіативність кореляції між ПФ та ПС ЗІК свідчить про суттєві відмінності широти охоплення супраспінальними впливами популяції мотонейронів ЗІК, їхнього виживання впродовж експерименту за різних видів нейротрансплантації.

The influence of neurotransplantation with different allogenic tissues on the course of the spasticity and chronic pain syndrome after experimental spinal cord injury

Objective. To examine the effect of neurotransplantation with different allogenic tissue types on the course of the spasticity and chronic pain syndrome after experimental spinal cord injury. Materials and methods. Animals: inbred albino male rats (5.5 months old,300 g); experimental groups: 1 — spinal cord injury + immediate homotopical transplantation of olfactory bulb tissue (TOBT, n = 34); 2 — spinal cord injury + analogous transplantation of fetal (E18) cerebellum tissue (TFСT, n = 15); 3 — spinal cord injury + analogous transplantation of fetal (E18) kidney tissue (TFKT, n = 8); 4 — spinal cord injury only (control group, n = 16). The model of injury was left-side spinal cord hemisection at Т11; spasticity in the ipsilateral hind limb was verified by Ashworth scale. Results. The increase (p 0.05). On the 24th week of observation the spasticity level in experimental groups was 2.6 ± 0.4 (control group), 2.2 ± 0.2 (TOBT), 2.1 ± 0.3 (TFСT) and 1.9 ± 0.3 (TFKT). Fifty-nine percent of the animals in TOBT group had early spasticity debut with flexion-adduction localization in hip and knee and peripheral paresis (hypotonia/atony) at the ankle joint and high prevalence of severe neurogenic pain manifestation (45 %). Similar spastic localization was noted in 40 % of the animals in TFСT group (for 2nd month) and 25 % of the animals in TFKT group (during 1st — 2nd week). In the control group signs of long-term severe neurogenic pain was found in 19 % animals, in TOBT group — in 27 %, in TFСT group — in 6 % (1 animal), in TFKT group no pain signs observed; the incidence difference was not significant. Conclusion. Approved types of neurotransplantation exert significant influence on the course of spasticity syndrome; the mechanisms of influence related to the cellular structure, angiogenic and immunogenic properties of the grafts.

Transcriptional response of a novel HpCDPK1 kinase gene from Hippeastrum x hybr. to wounding and fungal infection

Calcium dependent protein kinases (CDPK) are well established plant sensor and effectors for calcium ions and participate in regulation of multiple abiotic and biotic stress responses in plant cells. Here we present the identification and characterization of a new CDPK kinase gene from bulbous plant Hippeastrum x hybr. and examine the role of this kinase in stress responses leading to phytoalexin (PA) production in plant tissues. In the previous research, it was shown that Hippeastrum bulbs mechanically wounded or infected with Peyronellaea curtisii (=Phoma narcissi) are inducted to an antifungal red substance synthesis. In this research, we demonstrated Ca2+ dependence of the phytoalexin production by wounded bulbs. Furthermore, the isolated HpCDPK1 cDNA for ORF was found to be 1596bp long and encoded 531 amino acid protein with CDPK kinase activity, as was shown by recombinant GST-HpCDPK1 enzyme production and analysis. HpCDPK1 transcript was present in all vegetative and chosen generative organs of Hippeastrum plant. The dynamics of the observed HpCDPK1 mRNA changes in bulbs depended on stressor type. The mechanical injury caused one wave of transcript increase while more complex transcript changes were observed within 48h after Peyronellaea inoculation. In plant bulbs already accumulating red phytoalexin, increases in HpCDPK1 mRNA level were observed at certain intervals within 48h whereas, in the case of fungal infection, only one big increment in the transcript amount at the 10th minute after inoculation was detected. The observed transcriptional response of HpCDPK1 gene to wounding and pathogen infection stress suggests a positive correlation with phytoalexin synthesis and maintenance in bulb tissues and puts more light on CDPK kinase role in the plant stress response regulation. This also bears some potential for understanding the mechanism of a phytoalexin formation.

The effect of neurotransplantation of various allogeneic tissue types to motor function restore after experimental spinal cord injury

Objective. To examine the effect of different tissue type of neurotransplantation on the locomotor function restoration after experimental spinal cord injury. Materials and methods. Animals: inbred albino male rats ( 5.5 months, 300 g ); experimental groups: 1 — spinal cord injury + immediate homotopical transplantation of olfactory bulb tissue (TOBT, n=34), 2 — spinal cord injury + analogous transplantation of fetal (E18) cerebellum tissue (TFСT, n=15), 3 — spinal cord injury + analogous transplantation of fetal (E18) kidney tissue (TFKT, n=8 ), 4 — spinal cord injury only in similar (control–1, n=16) and different (control–2, n=40) experimental seasons. Model of injury — left-side spinal cord hemisection at Т XI level; monitoring the ipsilateral hindlimb function indicator (IHL FI) — the Вasso–Вeattie–Вresnahan scale (BBB). Results. The predominance (p> 0.05) of the IHL FI after approved types of neurotransplantation has been noted when comparing with the control group–1 — at the 1 st –5 th week (ТОBT), 1 st –2 nd and 6 th –7 th week (TFСT), and at the end of the 8 th week (TFKT); when comparing with the control group–2 — at the 1 st –3 rd (ТОBT) and 1 st (TFСT) week of the experiment. The maximum value of the IHL FI has been observed at the 2 nd (ТОBT, 3,7±0,5 points ВВВ), 1 st , 6 th –7 th (TFСT, 3,6±0,8 points ВВВ), 12 th and 20 th (TFKT, 3,6±1,2 points ВВВ) weeks, minimum value of the IHL FI — at the 24 th (ТОBT, 2,4±0,6 points ВВВ), 3 rd (TFСT, 3,0±0,9 points ВВВ) and 1 st (TFKT, 1,9±1,1 points ВВВ) week of the experiment. Average IHL FI values of the three experimental groups at the 24 th week of the experiment have been amounted to 2,4–3,3 points BBB and comprised in a range of control groups final mean values (1,6–3,4 балла ВВВ). Significant differences between the IHL FI values of the groups ТОBT, TFСT and TFKT have not been observed during the experiment. In the case of TOBT significant changes of IHL FI have been noted during the 2 nd (increase), 6 th –7 th and 16 th –24 th week (reduced to a level below the 1 st week); in the case of TFKT — at the 1 st –3 rd week (increase); in the case of TFСT no significant changes have been identified. A common feature of the dynamics of the three experimental groups is prevalence of IHL FI values over the control during the first few weeks and lack of progression during further period of observation, that can be interpreted in a view of angiogenic, neurotropic, proinflammatory and mediator effects of the grafts. Conclusion. Approved types of neurotransplantation provide a temporary effect, continuing during the first month of the traumatic process; the study of the pathophysiological mechanisms of this effect can significantly improve understanding of tissue processes after multicomponent neuroengineering interventions.

In vitro differentiation of neural stem cells derived from human olfactory bulb into dopaminergic-like neurons.

This study describes a new accessible source of neuronal stem cells that can be used in Parkinson's disease cell transplant. The human olfactory bulb contains neural stem cells (NSCs) that are responsible for neurogenesis in the brain and the replacement of damaged cellular components throughout life. NSCs are capable of differentiating into neuronal and glial cells. We isolated NSCs from the olfactory bulb of brain-death donors and differentiated them into dopaminergic neurons. The olfactory bulb tissues obtained were cultured in Dulbecco's modified Eagle's medium/nutrient mixture F12, B27 supplemented with basic fibroblast growth factor, epidermal growth factor and leukemia inhibitory factor. The NSCs and proliferation markers were assessed. The multipotentiality of olfactory bulb NSCs was demonstrated by their capacity to differentiate into neurons, oligodendrocytes and astrocytes. To generate dopaminergic neurons, olfactory bulb NSCs were differentiated in neurobasal medium, supplemented with B27, and treated with sonic hedgehog, fibroblast growth factor 8 and glial cell-derived neurotrophic factor from the 7th to the 21st day, followed by detection of dopaminergic neuronal markers including tyrosine hydroxylase and aromatic l-amino acid decarboxylase. The cells were expanded, established in continuous cell lines and differentiated into the two classical neuronal phenotypes. The percentage of co-positive cells (microtubule-associated protein 2 and tyrosine hydroxylase; aromatic l-amino acid decarboxylase and tyrosine hydroxylase) in the treated cells was significantly higher than in the untreated cells. These results illustrate the existence of multipotent NSCs in the adult human olfactory bulb that are capable of differentiating toward putative dopaminergic neurons in the presence of trophic factors. Taken together, our data encourage further investigations of the possible use of olfactory bulb NSCs as a promising cell-based therapeutic strategy for Parkinson's disease.

Fractionated low-dose exposure to ionizing radiation leads to DNA damage, epigenetic dysregulation, and behavioral impairment.

Studies of Fractionated Exposure to Low Doses of Ionizing Radiation (FELDIR) has become of increasing importance to clinical interventions. Its consequences on DNA damage, physical, and mental health have been insufficiently investigated, however. The goal of this study was to determine the effects of FELDIR on the brain using a mouse model. We addressed the levels of DNA damage, global genomic methylation, and DNA methylation machinery in cerebellum, frontal lobe, olfactory bulb and hippocampal tissues, as well as behavioral changes linked to FELDIR exposure. The results reveal increased levels of DNA damage, as reflected by increased occurrence of DNA Strand Breaks (SBs) and dysregulation of stress-response kinase p38. FELDIR also resulted in initial loss of global genomic methylation and altered expression of methyltransferases DNMT1 (down-regulation) and DNMT3a (up-regulation), as well as methyl-binding protein MeCP2 (up-regulation). FELDIR-associated behavioral changes included impaired skilled limb placement on a ladder rung task, increased rearing activity in an open field, and elevated anxiety-like behaviors. The said alterations showed significant dose and tissue specificity. Thus, FELDIR represents a critical impact on DNA integrity and behavioral outcomes that need to be considered in the design of clinical intervention studies.

Endurance assessment of Eichhornia crassipes (Mart.) Solms, in heavy metal contaminated site–A case study

AbstractIn this study, the ability to hyper accumulate heavy metals from contaminated site by water hyacinth (Eichhornia crassipes) was monitored. The heavy metal-contaminated habitat was validated by X-ray diffraction and energy dispersive X-ray elemental spectrometry analysis of the soil samples. Heavy metal contamination in soil and water; accumulation in foliar, root and bulb tissue samples were determined by atomic absorption spectroscopy and were monitored as a function of accumulation in different tissues. Significant differences were recorded in the bioaccumulation capability of heavy metals by different tissue. Discrete variation in protein profile of leaves and high expression of alcohol dehydrogenase (ADH), peroxidase (POX) and altered regulation of esterase (EST) in root tissue was observed in contaminated site grown Eichhornia. The high metal accumulation efficiency of water hyacinth due to the biomass production suggests this species as reliable organic biomarker for heavy metal contamination.

English

This study examined the phyto-toxic effects of a commonly named non-selective herbicide glyphosate (Roundup™) on onions (Allium cepa Linn.). The study was necessitated due to the indiscriminate use and release of Roundup™ for weed control in the Niger Delta soils of Nigeria. The Organisation for Economic Co-operation and Development (OECD) standard protocol # 208 was adopted. The short-term phyto-toxic consequence on onion (A. cepa L.) was determined after a 4-day exposure to varying concentrations of the test chemical at 0.625, 1.25, 2.5, 5 and 10 mg/L, respectively. The percentage growth rate decreased as percentage growth rate inhibition efficiency increased, which implies that the effect of the herbicide was concentration dependent. The mean percentage growth rate inhibition efficiency relative to the control was 28, 47, 61, 73 and 96%, respectively. The effective concentration (EC50) for % Inhibition efficiency was 1.550 mg/L with a 95% confidence interval of 1.269 to 1.848 mg/L. The biological alterations on the onions in the test solutions varied in order of increasing concentrations. Some observations made include decolouration of the test solutions and stunted growth especially at the highest concentration of 10 mg/L. Other effects include: bulb deformation, tissue and root damage. There was significant difference between the exposed species and the control at P < 0.05. The use of the herbicide Roundup™ with such hazardous effects can harm plants especially onion which is a very viable food product of man. This could lead to likely distortion on the ecosystem balance of onions (A. cepa) and similar plants and subsequent effect on human, the major end user. Key words: Herbicide, glyphosate (roundup), onion (Allium cepa Linn.), phytotoxicity.

The Effect of Chronic Methamphetamine Exposure on the Hippocampal and Olfactory Bulb Neuroproteomes of Rats.

Nowadays, drug abuse and addiction are serious public health problems in the USA. Methamphetamine (METH) is one of the most abused drugs and is known to cause brain damage after repeated exposure. In this paper, we conducted a neuroproteomic study to evaluate METH-induced brain protein dynamics, following a two-week chronic regimen of an escalating dose of METH exposure. Proteins were extracted from rat brain hippocampal and olfactory bulb tissues and subjected to liquid chromatography-mass spectrometry (LC-MS/MS) analysis. Both shotgun and targeted proteomic analysis were performed. Protein quantification was initially based on comparing the spectral counts between METH exposed animals and their control counterparts. Quantitative differences were further confirmed through multiple reaction monitoring (MRM) LC-MS/MS experiments. According to the quantitative results, the expression of 18 proteins (11 in the hippocampus and 7 in the olfactory bulb) underwent a significant alteration as a result of exposing rats to METH. 13 of these proteins were up-regulated after METH exposure while 5 were down-regulated. The altered proteins belonging to different structural and functional families were involved in processes such as cell death, inflammation, oxidation, and apoptosis.

Reactivation of HSV-1 following explant of tree shrew brain.

Herpes Simplex Virus type I (HSV-1) latently infects peripheral nervous system (PNS) sensory neurons, and its reactivation leads to recurring cold sores. The reactivated HSV-1 can travel retrograde from the PNS into the central nervous system (CNS) and is known to be causative of Herpes Simplex viral encephalitis. HSV-1 infection in the PNS is well documented, but little is known on the fate of HSV-1 once it enters the CNS. In the murine model, HSV-1 genome persists in the CNS once infected through an ocular route. To gain more details of HSV-1 infection in the CNS, we characterized HSV-1 infection of the tree shrew (Tupaia belangeri chinensis) brain following ocular inoculation. Here, we report that HSV-1 enters the tree shrew brain following ocular inoculation and HSV-1 transcripts, ICP0, ICP4, and LAT can be detected at 5 days post-infection (p.i.), peaking at 10 days p.i. After 2 weeks, ICP4 and ICP0 transcripts are reduced to a basal level, but the LAT intron region continues to be expressed. Live virus could be recovered from the olfactory bulb and brain stem tissue. Viral proteins could be detected using anti-HSV-1 antibodies and anti-ICP4 antibody, during the acute stage but not beyond. In situ hybridization could detect LAT during acute infection in most brain regions and in olfactory bulb and brain stem tissue well beyond the acute stage. Using a homogenate from these tissues’ post-acute infection, we did not recover live HSV-1 virus, supporting a latent infection, but using a modified explant cocultivation technique, we were able to recover reactivated virus from these tissues, suggesting that the HSV-1 virus latently infects the tree shrew CNS. Compared to mouse, the CNS acute infection of the tree shrew is delayed and the olfactory bulb contains most latent virus. During the acute stage, a portion of the infected tree shrews exhibit symptoms similar to human viral encephalitis. These findings, together with the fact that tree shrews are closely related to primates, provided a valuable alternative model to study HSV-1 infection and pathogenesis in the CNS.

Topical sulfonylurea as a novel therapy for hypertrichosis secondary to diazoxide, and potentially for other conditions with excess hair growth

It is hypothesized that a topical application of a sulfonylurea drug, which can inhibit the ATP-sensitive potassium-gated channels (Kir6.X/SUR) present in human hair bulb tissues, will inhibit hair growth in a targeted manner. Diazoxide is used to treat severe hypoglycemia due to hyperinsulinism of infancy. However, this often results in hypertrichosis that can be severe enough to prevent its use. Diazoxide blocks insulin release from the pancreas by opening the SUR1/Kir6.2 channels in ß-cells. Diazoxide can also act on two potassium-gated channels in the skin that affect hair growth, namely SUR1/Kir6.2 and SUR2B/Kir6.1, thus causing hypertrichosis. It is proposed that a topical sulfonylurea will inhibit the excessive hair growth due to diazoxide, but will not impact the beneficial effects of diazoxide on beta cells. This approach can also be applied to rare cases of Cantú syndrome, caused by mutations in ABCC9 (coding for SUR2) or in KCNJ8 (coding for Kir6.1) that is characterized by congenital hypertrichosis. More importantly, this approach may also be effective in treating other forms of hypertrichosis or hirsutism, that are quite common, yet very distressing to patients worldwide.

Kinetin Concentration Affect Somatic Embryogenic Callus by Bulb tissue of Onion(Allium cepa L.)

Kinetin Concentration Affect Somatic Embryogenic Callus by Bulb tissue of Onion(Allium cepa L.)

De Novo Transcriptome Analysis of Allium cepa L. (Onion) Bulb to Identify Allergens and Epitopes.

Allium cepa (onion) is a diploid plant with one of the largest nuclear genomes among all diploids. Onion is an example of an under-researched crop which has a complex heterozygous genome. There are no allergenic proteins and genomic data available for onions. This study was conducted to establish a transcriptome catalogue of onion bulb that will enable us to study onion related genes involved in medicinal use and allergies. Transcriptome dataset generated from onion bulb using the Illumina HiSeq 2000 technology showed a total of 99,074,309 high quality raw reads (~20 Gb). Based on sequence homology onion genes were categorized into 49 different functional groups. Most of the genes however, were classified under 'unknown' in all three gene ontology categories. Of the categorized genes, 61.2% showed metabolic functions followed by cellular components such as binding, cellular processes; catalytic activity and cell part. With BLASTx top hit analysis, a total of 2,511 homologous allergenic sequences were found, which had 37–100% similarity with 46 different types of allergens existing in the database. From the 46 contigs or allergens, 521 B-cell linear epitopes were identified using BepiPred linear epitope prediction tool. This is the first comprehensive insight into the transcriptome of onion bulb tissue using the NGS technology, which can be used to map IgE epitopes and prediction of structures and functions of various proteins.

Olfactory bulb as an alternative in neurotransplantation

The article examines the ethical and legal aspects of transplantation of embryonic neural tissue, structure of the rat olfactory bulb. It is given substantiation for its use as a possible alternative version of the embryonic neural tissue at damage in the cerebral hemispheres in the experiment.Materials and methods. Detailed description of the fault model of the cerebral hemispheres of the brain of rats, olfactory bulb biopsy procedure, cultivation of olfactory bulb suspension and fetal neural tissue, comparison of the functional aspects of transplantation of the olfactory bulb and the embryonic neural tissue.Results. The obtained data are similar to structure of olfactory bulb and fetal tissues during culturing. Recovery in the motor areas varies by the time factor and less intense in the group of the olfactory bulb and the group without tissue transplantation.Conclusions. Comparative analysis of the effectiveness of transplantation of embryonic neural tissue and olfactory bulb in the injured brain allows us to speak about the positive results of these groups to the difference in the duration of the recovery process

Hexacyclopeptides secreted by an endophytic fungus Fusarium solani N06 act as crosstalk molecules in Narcissus tazetta.

The basis of chemical crosstalk in plants and associated endophytes lies in certain so-called communication molecules that are responsible for plant-microbe and microbe-microbe interactions. Consequently, elucidating the factors that affect the nature, distribution, and amount of these molecules and how they impact the interaction among endophytes and associated organisms is essential to understand the true potential of endophytes. In the present study, we report the discovery of nine hexacyclopeptides from an endophytic fungus, Fusarium solani, isolated from the bulb of Narcissus tazetta, and their selective accumulation by an endophytic bacterium, Achromobacter xylosoxidans isolated from the same tissue. We used matrix-assisted laser desorption ionization imaging high-resolution mass spectrometry (MALDI-imaging-HRMS) to firstly identify and visualize the spatial distribution of the hexacyclopeptides produced by endophytic F. solani. After culture condition optimization, their sequence was identified to be cyclo((Hyp or Dhp)-Xle-Xle-(Ala or Val)-Thr-Xle) (Dhp: dehydroproline) by the characteristic a, b, or y ions using liquid chromatography tandem mass spectrometry (LC-HRMS(n)). These hexacyclopeptides were confirmed to be fungal biosynthetic products by deuterium labeling experiments. Finally, in order to understand the plausible ecological relevance of one or more of the discovered hexacyclopeptides within the contexts of microbial "neighbor communication," we devised a dual-culture setup to visualize using MALDI-imaging-HRMS how the hexacyclopeptides released by the endophytic fungus are accumulated by another endophytic bacterium, A. xylosoxidans, isolated from the same bulb tissue. This work exemplifies the relevance of cyclopeptides in endophyte-endophyte interspecies neighbor communication occurring in nature. Such communication strategies are evolved by coexisting endophytes to survive and function in their distinct ecological niches.

Microencapsulation improves inhibitory effects of transplanted olfactory ensheathing cells on pain after sciatic nerve injury.

Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplanted olfactory ensheathing cells (OECs) remains unclear. In the present study, we microencapsulated OECs in alginic acid, and transplanted free and microencapsulated OECs into the region surrounding the injured sciatic nerve in rat models of chronic constriction injury. We assessed mechanical nociception in the rat models 7 and 14 days after surgery by measuring paw withdrawal threshold, and examined P2X2/3 receptor expression in L4–5 dorsal root ganglia using immunohistochemistry. Rats that received free and microencapsulated OEC transplants showed greater withdrawal thresholds than untreated model rats, and weaker P2X2/3 receptor immunoreactivity in dorsal root ganglia. At 14 days, paw withdrawal threshold was much higher in the microencapsulated OEC-treated animals. Our results confirm that microencapsulated OEC transplantation suppresses P2X2/3 receptor expression in L4–5 dorsal root ganglia in rat models of neuropathic pain and reduces allodynia, and also suggest that transplantation of microencapsulated OECs is more effective than transplantation of free OECs for the treatment of neuropathic pain.

Detection of bacterial antigens and Alzheimer's disease-like pathology in the central nervous system of BALB/c mice following intranasal infection with a laboratory isolate of Chlamydia pneumoniae.

Pathology consistent with that observed in Alzheimer’s disease (AD) has previously been documented following intranasal infection of normal wild-type mice with Chlamydia pneumoniae (Cpn) isolated from an AD brain (96-41). In the current study, BALB/c mice were intranasally infected with a laboratory strain of Cpn, AR-39, and brain and olfactory bulbs were obtained at 1–4 months post-infection (pi). Immunohistochemistry for amyloid beta or Cpn antigens was performed on sections from brains of infected or mock-infected mice. Chlamydia-specific immunolabeling was identified in olfactory bulb tissues and in cerebrum of AR-39 infected mice. The Cpn specific labeling was most prominent at 1 month pi and the greatest burden of amyloid deposition was noted at 2 months pi, whereas both decreased at 3 and 4 months. Viable Cpn was recovered from olfactory bulbs of 3 of 3 experimentally infected mice at 1 and 3 months pi, and in 2 of 3 mice at 4 months pi. In contrast, in cortical tissues of infected mice at 1 and 4 months pi no viable organism was obtained. At 3 months pi, only 1 of 3 mice had a measurable burden of viable Cpn from the cortical tissues. Mock-infected mice (0 of 3) had no detectable Cpn in either olfactory bulbs or cortical tissues. These data indicate that the AR-39 isolate of Cpn establishes a limited infection predominantly in the olfactory bulbs of BALB/c mice. Although infection with the laboratory strain of Cpn promotes deposition of amyloid beta, this appears to resolve following reduction of the Cpn antigen burden over time. Our data suggest that infection with the AR-39 laboratory isolate of Cpn results in a different course of amyloid beta deposition and ultimate resolution than that observed following infection with the human AD-brain Cpn isolate, 96-41. These data further support that there may be differences, possibly in virulence factors, between Cpn isolates in the generation of sustainable AD pathology.

Comparison Study on Soil Physical and Chemical Properties, Plant Growth, Yield, and Nutrient Uptakes in Bulb Onion from Organic and Conventional Systems

This study aimed to compare agricultural practices, soil physical and chemical properties, growth characteristics, and nutrient uptake of bulb onions from organic and conventional farms in southeastern Korea during the 2011–12 growing season. Soil and plant samples were collected from eight certified organic fields managed organically for more than 5 years and eight conventional fields adjacent to the organic fields. The amounts of nutrients applied to onion fields were approximately two times greater with the conventional methods than with organic methods. However, the soil physical and chemical properties were not significantly different between the organic and conventional systems, except for NO3-N in early May. Growth characteristics were significantly different in early April with organic bulb yield of 55.9 t·ha−1, which was 21.8% lower than conventionally produced onions. Yield reduction in organic onions was the result of lower large-sized (8 cm or greater) bulb yield compared with conventional production. In the conventional system, phosphorus (P) and potassium (K) content of leaf tissue in early April, and nitrogen (N) and P content of bulb tissue in early May were higher than those in the organic system. Uptake of all nutrients was greater in the conventional onions compared with the organic onions, except for leaf tissue at harvest. In conclusion, organic onions began to grow and absorb soil nutrients later than the conventional onions in the initial vegetative growth stage. Moreover, it led to an organic onion producing a lower bulb yield. To accelerate the initial growth of the organic onion plant, agricultural practices need to be modified. Modifications that may help include using larger sized seedlings at transplanting, covering the plants with nonwoven fabric or transparent plastic film to increase warmth during winter, and harvesting the onions 1 week later than the conventional onions.

Combustion smoke-induced inflammation in the olfactory bulb of adult rats.

BackgroundThe damaging effect of combustion smoke inhalation on the lung is widely reported but information on its effects on the olfactory bulb is lacking. This study sought to determine the effects of smoke inhalation on the olfactory bulb, whose afferent input neurons in the nasal mucosa are directly exposed to external stimuli, such as smoke.MethodsAdult male Sprague-Dawley rats were subjected to combustion smoke inhalation and sacrificed at different time points. Changes in olfactory bulb proteins including vascular endothelial growth factor (VEGF), inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS), Na+-K+-Cl− cotransporter 1 (NKCC1), glial fibrillary acidic protein (GFAP), and aquaporin-4 (AQP4) were evaluated by Western blot analysis. In addition, ELISA was conducted for cytokine and chemokine levels, and double immunofluorescence labeling was carried out for GFAP/VEGF, GFAP/AQP4, NeuN/nNOS, GFAP/NKCC1, NeuN/NKCC1, GFAP/Rhodamine isothiocyanate (RITC), and transferase dUTP nick end labeling (TUNEL). Aminoguanidine was administered to determine the effects of iNOS inhibition on the targets probed after smoke inhalation.ResultsThe results showed a significant increase in VEGF, iNOS, eNOS, nNOS, NKCC1, and GFAP expression in the bulb tissues, with corresponding increases in inflammatory cytokines and chemokines after smoke inhalation. Concurrent to this was a drastic increase in AQP4 expression and RITC permeability. Aminoguanidine administration decreased the expression of iNOS and RITC extravasation after smoke inhalation. This was coupled with a significant reduction in incidence of TUNEL + cells that was not altered with administration of L-NG-nitroarginine methyl ester (L-NAME).ConclusionsThese findings suggest that the upregulation of iNOS in response to smoke inhalation plays a major role in the olfactory bulb inflammatory pathophysiology, along with a concomitant increase in pro-inflammatory molecules, vascular permeability, and edema. Overall, these findings indicate that the olfactory bulb is vulnerable to smoke inhalation.