Exploring methods of preparing functional cartilage-bone xenografts for joint repair

This thesis explores the feasibility of donor-receiver concept for joint replacement where cartilage-bone tissues can be taken from either human or other mammals and prepared scientifically for repairing focal joint defects in knees, hips and shoulders. The manufactured construct is immunologically inert and is capable of acting as a scaffold for engineering new cartilage-bone laminates when placed in the joint. Innovative manufacturing procedures and assessment techniques were developed for appraising this tissue-based scaffold. This research has demonstrated that tissue replacement technology can be applied in situations where blood vessels are absent such as in articular cartilage.

Amelioration of soil compaction can take 5 years on a Vertisol under no till in the semi-arid subtropics

Heavy wheel traffic causes soil compaction, which adversely affects crop production and may persist for several years. We applied known compaction forces to entire plots annually for 5 years, and then determined the duration of the adverse effects on the properties of a Vertisol and the performance of crops under no-till dryland cropping with residue retention. For up to 5 years after a final treatment with a 10 Mg axle load on wet soil, soil shear strength at 70-100 mm and cone index at 180-360 mm were significantly (P < 0.05) higher than in a control treatment, and soil water storage and grain yield were lower. We conclude that compaction effects persisted because (1) there were insufficient wet-dry cycles to swell and shrink the entire compacted layer, (2) soil loosening by tillage was absent and (3) there were fewer earthworms in the compacted soil. Compaction of dry soil with 6 Mg had little effect at any time, indicating that by using wheel traffic only when the soil is dry, problems can be avoided. Unfortunately such a restriction is not always possible because sowing, tillage and harvest operations often need to be done when the soil is wet. A more generally applicable solution, which also ensures timely operations, is the permanent separation of wheel zones and crop zones in the field--the practice known as controlled traffic farming. Where a compacted layer already exists, even on a clay soil, management options to hasten repair should be considered, e.g. tillage, deep ripping, sowing a ley pasture or sowing crop species more effective at repairing compacted soil.

Atomistic investigation into the mechanical behaviour of crystalline and amorphous TiO2 nanotubes

Titanium dioxide (TiO2) nanotubes are appealing to research communities due to their excellent functional properties. However, there is still a lack of understanding of their mechanical properties. In this work, we conduct molecular dynamics (MD) simulations to investigate the mechanical behaviour of rutile and amorphous TiO2 nanotubes. The results indicate that the rutile TiO2 nanotube has a much higher Young's modulus (∼800 GPa) than the amorphous one (∼400 GPa). Under tensile loading, rutile nanotubes fail in the form of brittle fracture but significant ductility (up to 30%) has been observed in amorphous nanotubes. This is attributed to a unique ‘repairing’ mechanism via bond reconstruction at under-coordinated sites as well as bond conversion at over-coordinated sites. In addition, it is observed that the fracture strength of rutile nanotubes is strongly dependent on their free surfaces. These findings are considered to be useful for development of TiO2 nanostructures with improved mechanical properties.

Cutaneous porphyrias : Clinical and histopathological study

The prevalence of variegate porphyria (VP) (2.1:100 000, in 2006 n=108) was higher in Finland than elsewhere in European countries due to a founder effect (R152C). The incidence of VP was estimated at 0.2:1 000 000 based on the number of new symptomatic patients yearly. The prevalence of porphyria cutanea tarda (PCT) was 1.2:100 000 (in 2006 n=63), which is only one fourth of the numbers reported from other European countries. The estimated incidence of PCT was 0.5:1 000 000. Based on measurements of the uroporphyrinogen decarboxylase activity in erythrocytes, the proportion of familial PCT was 49% of the cases. The prevalence of erythropoietic protoporphyria (EPP) was at 0.8:100 000 (in 2006 n=39) including asymptomatic carriers of a mutation in the ferrochelatase (FECH) gene. The incidence of EPP was estimated at 0.1:1 000 000. After 1980 the penetrance was 37% among patients with VP. Of the mutation carriers (n=57) 30% manifested with skin symptoms. Frequency of skin symptom as only clinical sign was stable before or after 1980 (22% vs. 21%), but acute attacks became infrequent (29% vs. 7%). Of the symptomatic patients 30% had both acute attacks and skin symptoms and 80% had skin symptoms. Fragility (95%) and blistering (46%) of the skin in the backs of the hands were the most common skin symptoms. Transient correction of porphyrin metabolism using eight haem arginate infusions within five weeks had no effect on the skin symptoms in three of four patients with VP. In one case skin symptoms disappeared transiently. One patient with homozygous VP had severe photosensitivity since birth. Sensory polyneuropathy, glaucoma and renal failure developed during the 25-year follow-up without the presence of acute attacks. The I12T mutation was detected in both of his alleles in the protoporphyrinogen oxidase gene. Lack of skin symptoms and infrequency of acute attacks (1/9) in the patients with I12T mutation at the heterozygous stage indicate a mild phenotype (the penetrance 11%). Four mutations (751delGAGAA, 1122delT, C286T, C343T) in the FECH gene were characterised in four of 15 families with EPP. Burning pain (96%) and swelling (92%) of the sun-exposed skin were the major skin symptoms. Hepatopathy appeared in one of 25 symptomatic patients (4%). Clinical manifestations and associated factors of PCT were similar in the sporadic and familial types of PCT. The majority of the patients with PCT had one to three precipitating factors: alcohol intake (78%), mutations in hemochromatosis associated gene (50%), use of oestrogen (25% of women) and hepatitis B or C infections (25 %). Fatty liver disease (67%) and siderosis (67%) were commonly found in their liver biopsies. The major histopathological change of the sun-exposed skin in the patients with VP (n=20), EPP (n=8) and PCT (n=5) was thickening of the vessel walls of the upper dermis suggesting that the vessel wall is the primary site of the phototoxic reaction in each type of porphyria. The fine structure of the vessel walls was similar in VP, EPP and PCT consisting of the multilayered basement membrane and excess of finely granular substance between the layers which were surrounded by the band of homogenous material. EPP was characterised by amorphous perivascular deposits extending also to the extravascular space. In direct immunofluorescence study homogenous IgG deposits in the vessel walls of the upper dermis of the sun-exposed skin were demonstrated in each type of porphyria. In EPP the excess material around vessel walls consisted of other proteins such as serum amyloid protein, and kappa and lambda light chains in addition to the basement membrane constituents such as collagen IV and laminin. These results suggest that the alterations of the vessel walls are a consequence of the repeated damage and the repairing process in the vessel wall. The microscopic alterations could be demonstrated even in the normal looking but sun-exposed skin of the patients with EPP during the symptom-free phase suggesting that vascular change can be chronic. The stability of vascular changes in the patients with PCT after treatment indicates that circulating porphyrins are not important for the maintenance of the changes.

Young love: Romantic concerns and associated mental health issues among adolescent help-seekers

Over 50% of young people have dated by age 15. While romantic relationship concerns are a major reason for adolescent help-seeking from counselling services, we have a limited understanding of what types of relationship issues are most strongly related to mental health issues and suicide risk. This paper used records of 4019 counselling sessions with adolescents (10–18 years) seeking help from a national youth counselling service for a romantic relationship concern to: (i) explore what types and stage (pre, during, post) of romantic concerns adolescents seek help for; (ii) how they are associated with mental health problems, self-harm and suicide risk; and (iii) whether these associations differ by age and gender. In line with developmental-contextual theory, results suggest that concerns about the initiation of relationships are common in early adolescence, while concerns about maintaining and repairing relationships increase with age. Relationship breakups were the most common concern for both male and female adolescents and for all age groups (early, mid, late adolescence). Data relating to a range of mental health issues were available for approximately half of the sample. Post-relationship concerns (including breakups) were also more likely than pre- or during-relationship concerns to be associated with concurrent mental health issues (36.8%), self-harm (22.6%) and suicide (9.9%). Results draw on a staged developmental theory of adolescent romantic relationships to provide a comprehensive assessment of relationship stressors, highlighting post-relationship as a particularly vulnerable time for all stages of adolescence. These findings contribute to the development of targeted intervention and support programs.

Nucleotide sequence of the first cosmid from the Mycobacterium leprae genome project: structure and function of the Rif-Str regions

The nucleotide sequence of cosmid B1790, carrying the Rif-Str regions of the Mycobacterium leprae chromosome, has been determined. Twelve open reading frames were identified in the 36716bp sequence, representing 40% of the coding capacity. Five ribosomal proteins, two elongation factors and the β and β'subunits of RNA polymerase have been characterized and two novel genes were found. One of these encodes a member of the so-called ABC family of ATP-binding proteins while the other appears to encode an enzyme involved in repairing genomic lesions caused by free radicals. This finding may well be significant as M. leprae, an intracellular pathogen, lives within macrophages.

Advances in the molecular design of potential anticancer agents via targeting of human telomeric DNA

Telomerases are an attractive drug target to develop new generation drugs against cancer. A telomere appears from the chromosomal termini and protects it from double-stranded DNA degradation. A short telomere promotes genomic instability, like end-to-end fusion and regulates the over-expression of the telomere repairing enzyme, telomerase. The telomerase maintains the telomere length, which may lead to genetically abnormal situations, leading to cancer. Thus, the design and synthesis of an efficient telomerase inhibitor is a viable strategy toward anticancer drugs development. Accordingly, small molecule induced stabilization of the G-quadruplex structure, formed by the human telomeric DNA, is an area of contemporary scientific art. Several such compounds efficiently stabilize the G-quadruplex forms of nucleic acids, which often leads to telomerase inhibition. This Feature article presents the discovery and development of the telomere structure, function and evolution in telomere targeted anticancer drug design and incorporates the recent advances in this area, in addition to discussing the advantages and disadvantages in the methods, and prospects for the future.

海洋平台用高强度钢多层补焊的疲劳分析

对海洋平台用高强度钢焊接接头多次补焊的疲劳性能进行试验研究，给出采用威布尔分布下的寿命估算式及R-S-N曲线。考虑到多次补焊后焊趾外移造成的焊接角的变化，应用有限元方法计算试样的应力分布，给出焊趾处的应力集中系数，由此进一步分析多次补焊对焊接接头疲劳性能的影响。

平台用高强度钢多次补焊及TIG熔修的疲劳分析

海洋石油平台在服役期内，由于受到各种环境载荷的交变作用，其受力的焊接构件不可避免地会出现疲劳裂纹。对出现的疲劳裂纹进行补焊修复，可以延长平台的使用期，带来巨大的经济和社会效益。本文通过实验研究，考察了多次补焊对海洋平台用高强度钢焊接接头疲劳性能的影响，以及是否可以通过TIG熔修工艺的采用提高补焊构件的疲劳寿命，达到延长补焊后平台的使用寿命，减少补焊次数的目的。本文涉及的主要研究内容如下：1、考察了多次补焊对焊接接头疲劳寿命的影响。通过对平台用高强度钢焊接接头初焊试样，以及重复三次补焊试样的疲劳实验，得到了各次补焊试样在对数正态分布和威布尔分布下的寿命估算式和考虑了存活率的R-S-N曲线。对实验结果进行了比较和分析，从而考察了多次补焊对焊接接头疲劳性能的影响。2、在每次补焊后，由于焊趾外移造成了焊趾角的变化，使得焊趾处的应力集中系数发生了变化。本文采用有限元方法计算了试样的应力场分布，得到了焊趾处的应力集中系数与焊趾角的关系。考虑到应力集中系数的变化后，对多次补焊试样疲劳实验的实验数据进行了重新处理。3、考察了多次补焊熔修对焊接接头疲劳寿命的影响。本文通过对平台用高强度钢焊接接头补焊及补焊熔修试样的疲劳实验，得到了多次补焊和补焊熔修试样对数正态分布和威布尔分布下的寿命估算式和考虑了存活率的R-S-N曲线。对实验结果进行了比较和分析，从而考察了补焊后采用TIG熔修工艺对焊接接头疲劳性能的影响。4、一次补焊后进行熔修处理可以极大的改善试样的疲劳性能，提高试样的疲劳寿命。但这一结果是在平焊位条件下得出了。为了将这一结果用于实际，还要考察实际操作中大量采用的立焊位条件下熔修的效果。本文完成了立焊位条件下补焊及补焊熔修试样的疲劳实验，得到了相应的S-N曲线。5、疲劳实验数据的处理中，大量采用威布尔分布作为数据统计的分布类型。但由于威布尔分布具有三个参数，给求解带来一定的困难。本文提出一种以在威布尔坐标纸上，数据分布的线形相关系数作为判据的求解威布尔参数的方法，并通过程序加以实现。为了对疲劳实验数据的处理更加方便和直观，编制了“疲劳实验数据处理程序“，将对疲劳实验数据的编辑、分布参数的求解、不同存活率下的疲劳寿命估算和各种分布类型图以及R-S-N曲线的绘制工作包括在该程序中。

Ca2+ Influx and Tyrosine Kinases Trigger Bordetella Adenylate Cyclase Toxin (ACT) Endocytosis. Cell Physiology and Expression of the CD11b/CD18 Integrin Major Determinants of the Entry Route

Humans infected with Bordetella pertussis, the whooping cough bacterium, show evidences of impaired host defenses. This pathogenic bacterium produces a unique adenylate cyclase toxin (ACT) which enters human phagocytes and catalyzes the unregulated formation of cAMP, hampering important bactericidal functions of these immune cells that eventually cause cell death by apoptosis and/or necrosis. Additionally, ACT permeabilizes cells through pore formation in the target cell membrane. Recently, we demonstrated that ACT is internalised into macrophages together with other membrane components, such as the integrin CD11b/CD18 (CR3), its receptor in these immune cells, and GM1. The goal of this study was to determine whether ACT uptake is restricted to receptor-bearing macrophages or on the contrary may also take place into cells devoid of receptor and gain more insights on the signalling involved. Here, we show that ACT is rapidly eliminated from the cell membrane of either CR3-positive as negative cells, though through different entry routes, which depends in part, on the target cell physiology and characteristics. ACT-induced Ca2+ influx and activation of non-receptor Tyr kinases into the target cell appear to be common master denominators in the different endocytic strategies activated by this toxin. Very importantly, we show that, upon incubation with ACT, target cells are capable of repairing the cell membrane, which suggests the mounting of an anti-toxin cell repair-response, very likely involving the toxin elimination from the cell surface.

Expanding the repertoire of DNA-mediated signaling in DNA repair

DNA damage is extremely detrimental to the cell and must be repaired to protect the genome. DNA is capable of conducting charge through the overlapping π-orbitals of stacked bases; this phenomenon is extremely sensitive to the integrity of the π-stack, as perturbations attenuate DNA charge transport (CT). Based on the E. coli base excision repair (BER) proteins EndoIII and MutY, it has recently been proposed that redox-active proteins containing metal clusters can utilize DNA CT to signal one another to locate sites of DNA damage.

To expand our repertoire of proteins that utilize DNA-mediated signaling, we measured the DNA-bound redox potential of the nucleotide excision repair (NER) helicase XPD from Sulfolobus acidocaldarius. A midpoint potential of 82 mV versus NHE was observed, resembling that of the previously reported BER proteins. The redox signal increases in intensity with ATP hydrolysis in only the WT protein and mutants that maintain ATPase activity and not for ATPase-deficient mutants. The signal increase correlates directly with ATP activity, suggesting that DNA-mediated signaling may play a general role in protein signaling. Several mutations in human XPD that lead to XP-related diseases have been identified; using SaXPD, we explored how these mutations, which are conserved in the thermophile, affect protein electrochemistry.

To further understand the electrochemical signaling of XPD, we studied the yeast S. cerevisiae Rad3 protein. ScRad3 mutants were incubated on a DNA-modified electrode and exhibited a similar redox potential to SaXPD. We developed a haploid strain of S. cerevisiae that allowed for easy manipulation of Rad3. In a survival assay, the ATPase- and helicase-deficient mutants show little survival, while the two disease-related mutants exhibit survival similar to WT. When both a WT and G47R (ATPase/helicase deficient) strain were challenged with different DNA damaging agents, both exhibited comparable survival in the presence of hydroxyurea, while with methyl methanesulfonate and camptothecin, the G47R strain exhibits a significant change in growth, suggesting that Rad3 is involved in repairing damage beyond traditional NER substrates. Together, these data expand our understanding of redox-active proteins at the interface of DNA repair.

Mitigating Scarring and Inflammation during Corneal Wound Healing using Nanofiber-Hydrogel Scaffolds

Due to the universal lack of donor tissue, there has been emerging interest in engineering materials to stimulate living cells to restore the features and functions of injured organs. We are particularly interested in developing materials for corneal use, where the necessity to maintain the tissue’s transparency presents an additional challenge. Every year, there are 1.5 – 2 million new cases of monocular blindness due to irregular healing of corneal injuries, dwarfing the approximately 150,000 corneal transplants performed. The large gap between the need and availability of cornea transplantation motivates us to develop a wound-healing scaffold that can prevent corneal blindness.

To develop such a scaffold, it is necessary to regulate the cells responsible for repairing the damaged cornea, namely myofibroblasts, which are responsible for the disordered and non-refractive index matched scar that leads to corneal blindness. Using in vitro assays, we identified that protein nanofibers of certain orientation can promote cell migration and modulate the myofibroblast phenotype. The nanofibers are also transparent, easy to handle and non-cytotoxic. To adhere the nanofibers to a wound bed, we examined the use of two different in situ forming hydrogels: an artificial extracellular matrix protein (aECM)-based gel and a photo-crosslinkable heparin-based gel. Both hydrogels can be formed within minutes, are transparent upon gelation and are easily tunable.

Using an in vivo mouse model for epithelial defects, we show that our corneal scaffolds (nanofibers together with hydrogel) are well-tolerated (no inflammatory response or turbidity) and support epithelium regrowth. We developed an ex vivo corneal tissue culture model where corneas that are wounded and treated with our scaffold can be cultured while retaining their ability to repair wounds for up to 21 days. Using this technique, we found that the aECM-based treatment induced a more favorable wound response than the heparin-based treatment, prompting us to further examine the efficacy of the aECM-based treatment in vivo using a rabbit model for stromal wounds. Results show that treated corneas have fewer myofibroblasts and immune cells than untreated ones, indicating that our corneal scaffold shows promise in promoting a calmer wound response and preventing corneal haze formation.

Produção literária: tecer o imaginário, espaço preenchido da margem

O objetivo deste estudo é analisar como a prática da produção literária interfere, positivamente, na trajetória de indivíduos que se encontram em situação de vulnerabilidade social. A investigação deu-se no Centro Cultural Cartola, situado na comunidade da Mangueira na cidade do Rio de Janeiro, espaço que favorece a experiência individual e social de crianças e jovens por meio de ferramentas pedagógico-culturais: dança, orquestra de violinos, música, capoeira, judô, balé, oficinas literárias de leituras e de composições de texto, que possibilitam a criação de valores culturais da comunidade e dos indivíduos pelo processo de identificação. A oficina Produção Literária: Tecer o Imaginário teve início em maio de 2011, com propostas de estímulo à criação e à leitura de textos ficcionais, possibilitando que a narrativa como um vetor de (re)construção do sujeito e do coletivo, sob o ponto de vista ético e estético. A fase investigativa constou de oficinas de literatura apresentadas de duas formas: Leitura e escrita em grupos pequenos - ou individuais. O trabalho de campo da nossa pesquisa qualitativo-participativa requereu que fizéssemos um recorte espacial de nossa experiência, para que à partir dai pudéssemos inserir o recorte teórico que fosse factível ao nosso objeto de investigação. Importava a dinâmica da ação dos atores sociais. A pesquisa, estruturada sobre a ação, revelou-se detentora de inúmeras possibilidades onde o imaginário se apresenta. A interação por meio de falas que estimulam a imaginação e a reflexão exerce influência no grupo e no indivíduo na construção de si e de seu mundo. Como o objetivo da pesquisa não se adequava somente à prática oral, de contar e ouvir estórias, mas de estimular os atores à inventar e escrevê-las percebemos que este modo prevaleceu. Por exigência de que a criança dominasse de alguma forma essa modalidade, sobretudo porque o ato de escrever validava as histórias inventadas por elas e as dispostas por outras pessoas nos livros. Visto que oralidade pode dispersar a entrada da criança em seu mundo imaginário, tendo importância maior para crianças muito pequenas que não entraram ainda no mundo da linguagem. A maneira que conduzíamos a pesquisa era sempre no sentido de estimular o imaginário, capturando-o, e a partir disso inseri-lo no mundo da linguagem, da simbolização. Para Minayo (1992), O método é o próprio processo do desenvolvimento das coisas.... Nesse sentido, surgia, daquela prática, um texto/sujeito, cujas palavras eram um tronco donde se espalhavam uma infinidade de ideias criativas. O resultado da investigação constatou que a criação ficcional, estimula e desenvolve o imaginário e as representações simbólicas empobrecidas. Evidenciando o quanto é gratificante e prazeroso para crianças e jovens suas participações em atividade que fazem do campo educativo e terapêutico um campo de aventura. Observar o efeito reparador da literatura no individuo, e de consolo nos momentos de lutos e de crises de toda ordem foi evidente em situação fronteiriça, pelo uso do mundo interno dessa potente ferramenta psicopedagogica como fortalecimento psicoemocional

A bone remodelling model coupling micro-damage growth and repair by 3D BMU-activity.

Bone as most of living tissues is able, during its entire lifetime, to adapt its internal microstructure and subsequently its associated mechanical properties to its specific mechanical and physiological environment in a process commonly known as bone remodelling. Bone is therefore continuously renewed and micro-damage, accumulated by fatigue or creep, is removed minimizing the risk of fracture. Nevertheless, bone is not always able to repair itself completely. Actually, if bone repairing function is slower than micro-damage accumulation, a type of bone fracture, usually known as "stress fracture", can finally evolve. In this paper, we propose a bone remodelling continuous model able to simulate micro-damage growth and repair in a coupled way and able therefore to predict the occurrence of "stress fractures". The biological bone remodelling process is modelled in terms of equations that describe the activity of basic multicellular units. The predicted results show a good correspondence with experimental and clinical data. For example, in disuse, bone porosity increases until an equilibrium situation is achieved. In overloading, bone porosity decreases unless the damage rate is so high that causes resorption or "stress fracture".

The socio-economic and cultural structures of the fisherfolk communities

Fishing communities that have exploited the resource for generations constitute the main stakeholder groups in the fisheries of Lake Victoria. Several studies have examined Uganda's Lake Victoria fishing communities and characterised key stakeholders at community level over the last decade (SEDAWOG 1999a and b; Geheb 1997; FeSEP 1997; Kitakule 1991). The communities are made up of scattered settlements at the shores and on islands. The categories of people living in these communities include fishers who consist primarily of large numbers of male youths who provide labour to boat and gear owners. There are resident and non-resident fish traders who after securing their supplies at the beaches, depart for their market destinations. In addition, there are fish processors, mostly operating traditional and improved smoking kilns. Many other people, dealing in provisions and supplies also stay at the beaches, their activities depending on the level of fish catch. The fishing communities of Lake Victoria, Uganda, include auxiliary livelihood activities such as boat building, net repairing and transportation; bait supply and beachside kiosks, video halls and retail shop business. Other economic activities are brick making, charcoal burning/wood trade, farming and livestock keeping.