Search results for: wounding
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 13

Search results for: wounding

13 Impact of Insect-Feeding and Fire-Heating Wounding on Wood Properties of Lodgepole Pine

Authors: Estelle Arbellay, Lori D. Daniels, Shawn D. Mansfield, Alice S. Chang

Abstract:

Mountain pine beetle (MPB) outbreaks are currently devastating lodgepole pine forests in western North America, which are also widely disturbed by frequent wildfires. Both MPB and fire can leave scars on lodgepole pine trees, thereby diminishing their commercial value and possibly compromising their utilization in solid wood products. In order to fully exploit the affected resource, it is crucial to understand how wounding from these two disturbance agents impact wood properties. Moreover, previous research on lodgepole pine has focused solely on sound wood and stained wood resulting from the MPB-transmitted blue fungi. By means of a quantitative multi-proxy approach, we tested the hypotheses that (i) wounding (of either MPB or fire origin) caused significant changes in wood properties of lodgepole pine and that (ii) MPB-induced wound effects could differ from those induced by fire in type and magnitude. Pith-to-bark strips were extracted from 30 MPB scars and 30 fire scars. Strips were cut immediately adjacent to the wound margin and encompassed 12 rings from normal wood formed prior to wounding and 12 rings from wound wood formed after wounding. Wood properties evaluated within this 24-year window included ring width, relative wood density, cellulose crystallinity, fibre dimensions, and carbon and nitrogen concentrations. Methods used to measure these proxies at a (sub-)annual resolution included X-ray densitometry, X-ray diffraction, fibre quality analysis, and elemental analysis. Results showed a substantial growth release in wound wood compared to normal wood, as both earlywood and latewood width increased over a decade following wounding. Wound wood was also shown to have a significantly different latewood density than normal wood 4 years after wounding. Latewood density decreased in MPB scars while the opposite was true in fire scars. By contrast, earlywood density was presented only minor variations following wounding. Cellulose crystallinity decreased in wound wood compared to normal wood, being especially diminished in MPB scars the first year after wounding. Fibre dimensions also decreased following wounding. However, carbon and nitrogen concentrations did not substantially differ between wound wood and normal wood. Nevertheless, insect-feeding and fire-heating wounding were shown to significantly alter most wood properties of lodgepole pine, as demonstrated by the existence of several morphological anomalies in wound wood. MPB and fire generally elicited similar anomalies, with the major exception of latewood density. In addition to providing quantitative criteria for differentiating between biotic (MPB) and abiotic (fire) disturbances, this study provides the wood industry with fundamental information on the physiological response of lodgepole pine to wounding in order to evaluate the utilization of scarred trees in solid wood products.

Keywords: elemental analysis, fibre quality analysis, lodgepole pine, wood properties, wounding, X-ray densitometry, X-ray diffraction

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12 Changes in the Body Weight and Wound Contraction Rate Following Treatment with Piper betel Extract in Diabetic Wounds

Authors: Nurul Z. Sani, Amalina N. Ghazali, Azree Elmy, Lee C. Yuen, Zar C. Thent

Abstract:

Piper betel (P. betel) leaves is widely used in Asian countries for treating diabetes mellitus and its complication. In our previous study, we observed the positive effect of P.betel extract on diabetic wounds following 3 and 7 days of treatment. The aim of the present study was to determine the effect of P.betel leaves extract in the diabetic rats was observed in terms of body weight and wound contraction rates following 5 days of the treatment. Total 64 male Sprague-Dawley rats were used and the experimental rats received a single dose of 60mg/kg of Streptozotocin (STZ) injection, intraperitoneally. Four full thickness (6mm) cutaneous wounds were created on dorsum of each rat. The rats were divideid into (n=8): Non-treated Control (NC), Non-treated Diabetic (ND), diabetic treated with commercial cream (SN) and diabetic treated with 50mg/kg of P.betel extract (PB). The rats were sacrificed on day 0 and 5 post wounding. Significant increased in wound closure rate, body weight was observed in PB group compared to ND. Histological deterioration was restored in the P. betel extract treated wounds. It is concluded that topical application with P.betel extract for 5 days of post wounding offers positive scientific value in diabetic rats.

Keywords: diabetes, piper betel, wound healing, body weight, morphology

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11 On the Estimation of Crime Rate in the Southwest of Nigeria: Principal Component Analysis Approach

Authors: Kayode Balogun, Femi Ayoola

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Crime is at alarming rate in this part of world and there are many factors that are contributing to this antisocietal behaviour both among the youths and old. In this work, principal component analysis (PCA) was used as a tool to reduce the dimensionality and to really know those variables that were crime prone in the study region. Data were collected on twenty-eight crime variables from National Bureau of Statistics (NBS) databank for a period of fifteen years, while retaining as much of the information as possible. We use PCA in this study to know the number of major variables and contributors to the crime in the Southwest Nigeria. The results of our analysis revealed that there were eight principal variables have been retained using the Scree plot and Loading plot which implies an eight-equation solution will be appropriate for the data. The eight components explained 93.81% of the total variation in the data set. We also found that the highest and commonly committed crimes in the Southwestern Nigeria were: Assault, Grievous Harm and Wounding, theft/stealing, burglary, house breaking, false pretence, unlawful arms possession and breach of public peace.

Keywords: crime rates, data, Southwest Nigeria, principal component analysis, variables

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10 Combining Transcriptomics, Bioinformatics, Biosynthesis Networks and Chromatographic Analyses for Cotton Gossypium hirsutum L. Defense Volatiles Study

Authors: Ronald Villamar-Torres, Michael Staudt, Christopher Viot

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Cotton Gossypium hirsutum L. is one of the most important industrial crops, producing the world leading natural textile fiber, but is very prone to arthropod attacks that reduce crop yield and quality. Cotton cultivation, therefore, makes an outstanding use of chemical pesticides. In reaction to herbivorous arthropods, cotton plants nevertheless show natural defense reactions, in particular through volatile organic compounds (VOCs) emissions. These natural defense mechanisms are nowadays underutilized but have a very high potential for cotton cultivation, and elucidating their genetic bases will help to improve their use. Simulating herbivory attacks by mechanical wounding of cotton plants in greenhouse, we studied by qPCR the changes in gene expression for genes of the terpenoids biosynthesis pathway. Differentially expressed genes corresponded to higher levels of the terpenoids biosynthesis pathway and not to enzymes synthesizing particular terpenoids. The genes were mapped on the G. hirsutum L. reference genome; their global relationships inside the general metabolic pathways and the biosynthesis of secondary metabolites were visualized with iPath2. The chromatographic profiles of VOCs emissions indicated first monoterpenes and sesquiterpenes emissions, dominantly four molecules known to be involved in plant reactions to arthropod attacks. As a result, the study permitted to identify potential key genes for the emission of volatile terpenoids by cotton plants in reaction to an arthropod attack, opening possibilities for molecular-assisted cotton breeding in benefit of smallholder cotton growers.

Keywords: biosynthesis pathways, cotton, mechanisms of plant defense, terpenoids, volatile organic compounds

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9 Literature as a Strategic Tool to Conscientise Africans: An Attempt by Postcolonial Writers and Critics to Reverse the Socio-Economics Imbalances of Colonialism

Authors: Lutendo Nendauni

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Colonialism breaks things, colonisers exploded native cultural solidarity, producing the spiritual confusion, psychic wounding, and economic exploitation of a new and dominated ‘other’. Colonialism as the cultural and economic exploitation began when the West defended in their seizure of foreign territories for the exploitation of its natural resources; this resulted in brutal socio-economic imbalances. The Western profited at the detriment of the weak Africa. However, colonialism has since passed, but the effects are still evident culturally, socially, and economically. This paper explored how postcolonial writers and critics attempt to reverse the socio-economic imbalances resulting from the fragmentation of colonialism, with a focus on the play 'I will Marry When I Want' by Ngugi wa Thiong’o and Ngugi wa Mirii, as a primary text. Using qualitative discourse-textual analysis as the research methodology, the researcher purposively extracts discourse segments from the text for analysis and interpretation. The findings reveal that Postcolonial critics and writers attempt to reverse the socio-economic effects of colonialism through various counter discourses; their literature is concerned with the destruction of colonised identity, the search for this identity, and its assertion. It is manifest in the text that writers offer corrective views about Africans; they stress that they write their literary texts to conscientise their fellow Africans. Postcolonial writers and critics argue that language is a carrier of culture and that the only way to break free from colonial influence is by not adopting a foreign language. They further through their poems, novels, plays, and music strategically shine the spotlight on the previously nameless and destitute people so that they can develop the human spirit’s desire to overcome defeat, socio-political deprivation, and isolation.

Keywords: colonialism, postcoloniality, critics, socio-economic imbalances

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8 WILCKO-PERIO, Periodontally Accelerated Orthodontics

Authors: Kruttika Bhuse

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Aim: Synergism between periodontists and orthodontists (periodontal accelerated osteogenic orthodontics- PAOO) creates crucial opportunities to enhance clinical outcomes of combined therapies regarding both disciplines and has made adult orthodontics a reality. Thus, understanding the biomechanics of bone remodelling may increase the clinical applications of corticotomy facilitated orthodontics with or without alveolar augmentation. Wilckodontics can be an attractive treatment option and be a “win-win” situation for both the dental surgeon and patient by reducing the orthodontic treatment time in adults. Materials and methods: In this review, data related to the clinical aspects, steps of procedure, biomechanics of bone, indications and contraindications and final outcome of wilckodontic shall be discussed. 50 supporting articles from various international journals and 70 clinical cases were reviewed to get a better understanding to design this wilckodontic - meta analysis. Various journals like the Journal Of Clinical And Diagnostic Research, Journal Of Indian Society Of Periodontology, Journal Of Periodontology, Pubmed, Boston Orthodontic University Journal, Good Practice Orthodontics Volume 2, have been referred to attain valuable information on wilckodontics which was then compiled in this single review study. Result: As a promising adjuvant technique based on the transient nature of demineralization-remineralisation process in healthy tissues, wilckodontics consists of regional acceleratory phenomenon by alveolar corticotomy and bone grafting of labial and palatal/lingual surfaces, followed by orthodontic force. The surgical wounding of alveolar bone potentiates tissue reorganization and healing by a way of transient burst of localized hard and soft tissue remodelling.This phenomenon causes bone healing to occur 10-50 times faster than normal bone turnover. Conclusion: This meta analysis helps understanding that the biomechanics of bone remodelling may increase the clinical applications of corticotomy facilitated orthodontics with or without alveolar augmentation. The main benefits being reduced orthodontic treatment time, increased bone volume and post-orthodontic stability.

Keywords: periodontal osteogenic accelerated orthodontics, alveolar corticotomy, bone augmentation, win-win situation

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7 De Novo Assembly and Characterization of the Transcriptome from the Fluoroacetate Producing Plant, Dichapetalum Cymosum

Authors: Selisha A. Sooklal, Phelelani Mpangase, Shaun Aron, Karl Rumbold

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Organically bound fluorine (C-F bond) is extremely rare in nature. Despite this, the first fluorinated secondary metabolite, fluoroacetate, was isolated from the plant Dichapetalum cymosum (commonly known as Gifblaar). However, the enzyme responsible for fluorination (fluorinase) in Gifblaar was never isolated and very little progress has been achieved in understanding this process in higher plants. Fluorinated compounds have vast applications in the pharmaceutical, agrochemical and fine chemicals industries. Consequently, an enzyme capable of catalysing a C-F bond has great potential as a biocatalyst in the industry considering that the field of fluorination is virtually synthetic. As with any biocatalyst, a range of these enzymes are required. Therefore, it is imperative to expand the exploration for novel fluorinases. This study aimed to gain molecular insights into secondary metabolite biosynthesis in Gifblaar using a high-throughput sequencing-based approach. Mechanical wounding studies were performed using Gifblaar leaf tissue in order to induce expression of the fluorinase. The transcriptome of the wounded and unwounded plant was then sequenced on the Illumina HiSeq platform. A total of 26.4 million short sequence reads were assembled into 77 845 transcripts using Trinity. Overall, 68.6 % of transcripts were annotated with gene identities using public databases (SwissProt, TrEMBL, GO, COG, Pfam, EC) with an E-value threshold of 1E-05. Sequences exhibited the greatest homology to the model plant, Arabidopsis thaliana (27 %). A total of 244 annotated transcripts were found to be differentially expressed between the wounded and unwounded plant. In addition, secondary metabolic pathways present in Gifblaar were successfully reconstructed using Pathway tools. Due to lack of genetic information for plant fluorinases, a transcript failed to be annotated as a fluorinating enzyme. Thus, a local database containing the 5 existing bacterial fluorinases was created. Fifteen transcripts having homology to partial regions of existing fluorinases were found. In efforts to obtain the full coding sequence of the Gifblaar fluorinase, primers were designed targeting the regions of homology and genome walking will be performed to amplify the unknown regions. This is the first genetic data available for Gifblaar. It has provided novel insights into the mechanisms of metabolite biosynthesis and will allow for the discovery of the first eukaryotic fluorinase.

Keywords: biocatalyst, fluorinase, gifblaar, transcriptome

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6 Lactobacillus rhamnosus GG Increases the Re-Epithelialization Rate of Model Wounds by Stimulating Keratinocyte Migration in Ex-Vivo

Authors: W. Mohammedsaeed, A. J. Mcbain, S. M. Cruickshank, C. A. O’Neill

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Many studies have demonstrated the importance of probiotics and their potential therapeutic effects within the gut. Recently, the possible therapeutic effects of probiotics in other tissues have also begun to be investigated. Comparatively few studies have evaluated the use of topical probiotics in relation to the skin. In this study, we have conducted preliminary investigations into whether a well-known probiotic, Lactobacillus rhamnosus GG (LGG), can increase the rate of re-epithelialization in a model wound. Full-thickness skin was obtained from individuals undergoing elective cosmetic surgery. This skin was wounded using excisional punch and cultured using a serum-free medium, either in the presence or absence of L. rhamnosus GG lysate. Histological staining of the sections was performed with Haematoxylin& Eosin E to quantify “epithelial tongue length”. This is the length of the new epithelial ‘tongue’ that grows and covers the exposed dermis at the inner wound edges. The length of the new epithelial ‘tongue’ was compared in untreated section and section treated with and L. rhamnosus GG made using108CFU/ml bacterial cells. L. rhamnosus GG lysate enhanced significantly the re-epithelialisation of treated wounds compared with that of untreated wounds (P=0.005, n=3). Tongue length, at day 1 was 7.55μm 0.15, at day 3 it was 18.5μm 0.25 and at day 7 was 22.9μm 0.35. These results can be compared with untreated cultures in which tongue length was 3.25μm 0.35, day 3 was 9.65μm 0.25 and day 7 was 13.5μm 0.15 post-wounding. In ex-vivo proliferation and migration cells were measured by determining the expression of nuclear proliferation marker Ki-67 and the expression of Phosphorylated cortactin respectively demonstrated that L. rhamnosus GG significantly increased NHEK proliferation and migration rates relative to controls. However, the dominant mechanism was migration because in ex-vivo skin treated with the L. rhamnosus GG up-regulated the gene expression of the chemokine receptor and ligands CXCR2 and CXCL2 comparing with controls (P=0.02, P=0.03 respectively, n=3). High levels of CXCL2/CXCL2 have already been implicated in multiple aspects of stimulation of wound healing through activation of keratinocyte migration. These data demonstrate that lysates from Lactobacillus rhamnosus GG increase re-epithelialization by stimulation of keratinocyte migration. The current study identifies the partial mechanism that contribute to stimulating the wound-healing process ex vivo in response to L. rhamnosus GG lysate is an increase in the production of CXCL2/ CXCR2 in ex vivo models. The use of probiotic lysates potentially offers new options to develop treatments that could improve wound healing.

Keywords: Lactobacillus rhamnosus GG, wounds, migration, lysate

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5 A Preliminary Study on the Effects of Lung Impact on Ballistic Thoracic Trauma

Authors: Amy Pullen, Samantha Rodrigues, David Kieser, Brian Shaw

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The aim of the study was to determine if a projectile interacting with the lungs increases the severity of injury in comparison to a projectile interacting with the ribs or intercostal muscle. This comparative study employed a 10% gelatine based model with either porcine ribs or balloons embedded to represent a lung. Four sample groups containing five samples were evaluated; these were control (plain gel), intercostal impact, rib impact, and lung impact. Two ammunition natures were evaluated at a range of 10m; these were 5.56x45mm and 7.62x51mm. Aspects of projectile behavior were quantified including exiting projectile weight, location of yawing, projectile fragmentation and distribution, location and area of the temporary cavity, permanent cavity formation, and overall energy deposition. Major findings included the cavity showing a higher percentage of the projectile weight exit the block than the intercostal and ribs, but similar to the control for the 5.56mm ammunition. However, for the 7.62mm ammunition, the lung was shown to have a higher percentage of the projectile weight exit the block than the control, intercostal and ribs. The total weight of projectile fragments as a function of penetration depth revealed large fluctuations and significant intra-group variation for both ammunition natures. Despite the lack of a clear trend, both plots show that the lung leads to greater projectile fragments exiting the model. The lung was shown to have a later center of the temporary cavity than the control, intercostal and ribs for both ammunition types. It was also shown to have a similar temporary cavity volume to the control, intercostal and ribs for the 5.56mm ammunition and a similar temporary cavity to the intercostal for the 7.62mm ammunition The lung was shown to leave a similar projectile tract than the control, intercostal and ribs for both ammunition types. It was also shown to have larger shear planes than the control and the intercostal, but similar to the ribs for the 5.56mm ammunition, whereas it was shown to have smaller shear planes than the control but similar shear planes to the intercostal and ribs for the 7.62mm ammunition. The lung was shown to have less energy deposited than the control, intercostal and ribs for both ammunition types. This comparative study provides insights into the influence of the lungs on thoracic gunshot trauma. It indicates that the lungs limits projectile deformation and causes a later onset of yawing and subsequently limits the energy deposited along the wound tract creating a deeper and smaller cavity. This suggests that lung impact creates an altered pattern of local energy deposition within the target which will affect the severity of trauma.

Keywords: ballistics, lung, trauma, wounding

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4 Relationship Demise After Having Children: An Analysis of Abandonment and Nuclear Family Structure vs. Supportive Community Cultures

Authors: John W. Travis

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There is an epidemic of couples separating after a child is born into a family, generally with the father leaving emotionally or physically in the first few years after birth. This separation creates high levels of stress for both parents, especially the primary parent, leaving her (or him) less available to the infant for healthy attachment and nurturing. The deterioration of the couple’s bond leaves parents increasingly under-resourced, and the dependent child in a compromised environment, with an increased likelihood of developing an attachment disorder. Objectives: To understand the dynamics of a couple, once the additional and extensive demands of a newborn are added to a nuclear family structure, and to identify effective ways to support all members of the family to thrive. Qualitative studies interviewed men, women, and couples after pregnancy and the early years as a family, regarding key destructive factors, as well as effective tools for the couple to retain a strong bond. In-depth analysis of a few cases, including the author’s own experience, reveal deeper insights about subtle factors, replicated in wider studies. Using a self-assessment survey, many fathers report feeling abandoned, due to the close bond of the mother-baby unit, and in turn, withdrawing themselves, leaving the mother without support and closeness to resource her for the baby. Fathers report various types of abandonment, from his partner to his mother, with whom he did not experience adequate connection as a child. The study identified a key destructive factor to be unrecognized wounding from childhood that was carried into the relationship. The study culminated in the naming of Male Postpartum Abandonment Syndrome (MPAS), describing the epidemic in industrialized cultures with the nuclear family as the primary configuration. A growing family system often collapses without a minimum number of adult caregivers per infant, approximately four per infant (3.87), which allows for proper healing and caretaking. In cases with no additional family or community beyond one or two parents, the layers of abandonment and trauma result in the deterioration of a couple’s relationship and ultimately the family structure. The solution includes engaging community in support of new families. The study identified (and recommends) specific resources to assist couples in recognizing and healing trauma and disconnection at multiple levels. Recommendations include wider awareness and availability of resources for healing childhood wounds and greater community-building efforts to support couples for the whole family to thrive.

Keywords: abandonment, attachment, community building, family and marital functioning, healing childhood wounds, infant wellness, intimacy, marital satisfaction, relationship quality, relationship satisfaction

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3 Characteristics of Female Offenders: Using Childhood Victimization Model for Treatment

Authors: Jane E. Hill

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Sexual, physical, or emotional abuses are behaviors used by one person in a relationship or within a family unit to control the other person. Physical abuse can consist of, but not limited to hitting, pushing, and shoving. Sexual abuse is unwanted or forced sexual activity on a person without their consent. Abusive behaviors include intimidation, manipulation, humiliation, isolation, frightening, terrorizing, coercing, threatening, blaming, hurting, injuring, or wounding another individual. Although emotional, psychological and financial abuses are not criminal behaviors, they are forms of abuse and can leave emotional scars on their victim. The purpose of this literature review research was to examine characteristics of female offenders, past abuse, and pathways to offending. The question that guided this research: does past abuse influence recidivism? The theoretical foundation used was relational theory by Jean Baker Miller. One common feature of female offenders is abuse (sexual, physical, or verbal). Abuse can cause mental illnesses and substance abuse. The abuse does not directly affect the women's recidivism. However, results indicated the psychological and maladaptive behaviors as a result of the abuse did contribute to indirect pathways to continue offending. The female offenders’ symptoms of ongoing depression, anxiety, and engaging in substance abuse (self medicating) did lead to the women's incarceration. Using the childhood victimization model as the treatment approach for women's mental illness and substance abuse disorders that were a result from history of child abuse have shown success. With that in mind, if issues surrounding early victimization are not addressed, then the women offenders may not recover from their mental illness or addiction and are at a higher risk of reoffending. However, if the women are not emotionally ready to engage in the treatment process, then it should not be forced onto them because it may cause harm (targeting prior traumatic experiences). Social capital is family support and sources that assist in helping the individual with education, employment opportunities that can lead to success. Human capital refers to internal knowledge, skills, and capacities that help the individual act in new and appropriate ways. The lack of human and social capital is common among female offenders, which leads to extreme poverty and economic marginalization, more often in frequent numbers than men. In addition, the changes in welfare reform have exacerbated women’s difficulties in gaining adequate-paying jobs to support themselves and their children that have contributed to female offenders reoffending. With that in mind, one way to lower the risk factor of female offenders from reoffending is to provide them with educational and vocational training, enhance their self-efficacy, and teach them appropriate coping skills and life skills. Furthermore, it is important to strengthen family bonds and support. Having a supportive family relationship was a statistically significant protective factor for women offenders.

Keywords: characteristics, childhood victimization model, female offenders, treatment

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2 Plant Regeneration via Somatic Embryogenesis and Agrobacterium-Mediated Transformation in Alfalfa (Medicago sativa L.)

Authors: Sarwan Dhir, Suma Basak, Dipika Parajulee

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Alfalfa is renowned for its nutritional and biopharmaceutical value as a perennial forage legume. However, establishing a rapid plant regeneration protocol using somatic embryogenesis and efficient transformation frequency are the crucial prerequisites for gene editing in alfalfa. This study was undertaken to establish and improve the protocol for somatic embryogenesis and subsequent plant regeneration. The experiments were conducted in response to natural sensitivity using various antibiotics such as cefotaxime, carbenicillin, gentamycin, hygromycin, and kanamycin. Using 3-week-old leaf tissue, somatic embryogenesis was initiated on Gamborg’s B5 basal (B5H) medium supplemented with 3% maltose, 0.9µM Kinetin, and 4.5µM 2,4-D. Embryogenic callus (EC) obtained from the B5H medium exhibited a high rate of somatic embryo formation (97.9%) after 3 weeks when the cultures were placed in the dark. Different developmental stages of somatic embryos and cotyledonary stages were then transferred to Murashige and Skoog’s (MS) basal medium under light, resulting in a 94% regeneration rate of plantlets. Our results indicate that leaf segments can grow (tolerate) up to 450 mg/L of cefotaxime and 400 mg/L of carbenicillin in the culture medium. However, the survival threshold for hygromycin at 12.5 mg/L, kanamycin at 250 mg/L, gentamycin at 50 mg/L, and timentin (300 mg/L). The experiment to improve the protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector was also conducted. The vector contains two reporter genes such as β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene (HPT), all driven under the CaMV 35s promoter. Various transformation parameters were optimized using 3-week-old in vitro-grown plantlets. The different parameters such as types of explant, leaf ages, preculture days, segment sizes, wounding types, bacterial concentrations, infection periods, co-cultivation periods, different concentrations of acetosyringone, silver nitrate, and calcium chloride were optimized for transient gene expression. The transient gene expression was confirmed via histochemical GUS and GFP visualization under fluorescent microscopy. The data were analyzed based on the semi-quantitative observation of the percentage and number of blue GUS spots on different days of agro-infection. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old leaf segments wounded using a scalpel blade of 11 size- after 3 days of post-incubation at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of bacterial-leaf segment co-cultivation, with the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. Our results suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The stable gene expression in the putative transgenic tissue was confirmed using PCR amplification of both marker genes, indicating that gene expression in explants was not solely due to Agrobacterium, but also from transformed cells. The improved protocol could be used for generating transgenic alfalfa plants using genome editing techniques such as CRISPR/Cas9.

Keywords: Medicago sativa l. (Alfalfa), agrobacterium tumefaciens, β-glucuronidase, green fluorescent protein, transient gene

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1 Heterotopic Ossification: DISH and Myositis Ossificans in Human Remains Identification

Authors: Patricia Shirley Almeida Prado, Liz Brito, Selma Paixão Argollo, Gracie Moreira, Leticia Matos Sobrinho

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Diffuse idiopathic skeletal hyperostosis (DISH) is a degenerative bone disease also known as Forestier´s disease and ankylosing hyperostosis of the spine is characterized by a tendency toward ossification of half the anterior longitudinal spinal ligament without intervertebral disc disease. DISH is not considered to be osteoarthritis, although the two conditions commonly occur together. Diagnostic criteria include fusion of at least four vertebrae by bony bridges arising from the anterolateral aspect of the vertebral bodies. These vertebral bodies have a 'dripping candle wax' appearance, also can be seen periosteal new bone formation on the anterior surface of the vertebral bodies and there is no ankylosis at zygoapophyseal facet joint. Clinically, patients with DISH tend to be asymptomatic some patients mention moderate pain and stiffness in upper back. This disease is more common in man, uncommon in patients younger than 50 years and rare in patients under 40 years old. In modern populations, DISH is found in association with obesity, (type II) diabetes; abnormal vitamin A metabolism and also associated with higher levels of serum uric acid. There is also some association between the increase of risk of stroke or other cerebrovascular disease. The DISH condition can be confused with Heterotopic Ossification, what is the bone formation in the soft tissues as the result of trauma, wounding, surgery, burnings, prolonged immobility and some central nervous system disorder. All these conditions have been described extensively as myositis ossificans which can be confused with the fibrodysplasia (myositis) ossificans progressive. As in the DISH symptomatology it can be asymptomatic or extensive enough to impair joint function. A third confusion osteoarthritis disease that can bring confusion are the enthesopathies that occur in the entire skeleton being common on the ischial tuberosities, iliac crests, patellae, and calcaneus. Ankylosis of the sacroiliac joint by bony bridges may also be found. CASE 1: this case is skeletal remains presenting skull, some vertebrae and scapulae. This case remains unidentified and due to lack of bone remains. Sex, age and ancestry profile was compromised, however the DISH pathognomonic findings and diagnostic helps to estimate sex and age characteristics. Moreover to presenting DISH these skeletal remains also showed some bone alterations and non-metrics as fusion of the first vertebrae with occipital bone, maxillae and palatine torus and scapular foramen on the right scapulae. CASE 2: this skeleton remains shows an extensive bone heterotopic ossification on the great trochanter area of left femur, right fibula showed a healed fracture in its body however in its inteosseous crest there is an extensive bone growth, also in the Ilium at the region of inferior gluteal line can be observed some pronounced bone growth and the skull presented a pronounced mandibular, maxillary and palatine torus. Despite all these pronounced heterotopic ossification the whole skeleton presents moderate bone overgrowth that is not linked with aging, since the skeleton belongs to a young unidentified individual. The appropriate osteopathological diagnosis support the human identification process through medical reports and also assist with epidemiological data that can strengthen vulnerable anthropological estimates.

Keywords: bone disease, DISH, human identification, human remains

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