Designing self sustaining support for micro-enterprises in sub-Saharan Africa : a pilot

Small and micro-enterprises play a significant part in most economies. However, in developing countries these enterprises have often been planned and implemented from a poverty alleviation perspective, rather than as entrepreneurial activities and growing businesses (Mead & Liedholm 1998, Toye 1993). This paper explores a model for sustainable support to micro-enterprises in a developing country context. Sustainability is critical if micro-enterprises are to grow. The traditional philanthropic model for funding micro-enterprises leaves local communities dependent on the priorities of donors which may not always be consistent with those of the community. Long term sustainability requires a move to a model that broadens the base of both economic and intellectual resources, that builds capacity as well as providing ‘start-up’ funding through mechanism such as micro-credit (Elkington & Hartigan 2008). Designing for sustainable enterprise development includes responding to the community’s priorities, investigating individual problems and solutions, encouraging agency and active involvement in goal setting, with on-going consultation and co-development of solutions. Recognising the importance of experimentation we have moved beyond existing forms to prototype new ways of working which provide continuity of financial and intellectual support for local initiatives. The paper reviews existing micro finance and microcredit practices and suggests a new approach to establish and support enterprises with financial and knowledge resources for sustainable business practices. Principles underlying such a program and the initial steps are described.

Teaching English : some remarks on the emergence of the sympathetic teacher in the English classroom

The chapter explores the concept of the teacher-student relationship within the English classroom and proposes that this relationship has a specific purpose in building a civil society by inculcating practices of self-reflection, empathy and ethics. The topic is explored through the example of teaching Australian literature, specifically, Tim Winton’s Lockie Leonard, Human Torpedo to secondary school students. Ian Hunter’s work on literature education and the construction of socio-ethical practices provides a conceptual framework for the chapter.

The Relational Self, the Social Bond and the Dynamics of Personal Relationships : A Sociological Analysis

This study analyses personal relationships linking research to sociological theory on the questions of the social bond and on the self as social. From the viewpoint of disruptive life events and experiences, such as loss, divorce and illness, it aims at understanding how selves are bound to their significant others as those specific people ‘close or otherwise important’ to them. Who form the configurations of significant others? How do different bonds respond in disruptions and how do relational processes unfold? How is the embeddedness of selves manifested in the processes of bonding, on the one hand, and in the relational formation of the self, on the other? The bonds are analyzed from an anti-categorical viewpoint based on personal citations of significance as opposed to given relationship categories, such as ‘family’ or ‘friendship’ – the two kinds of relationships that in fact are most frequently significant. The study draws from analysis of the personal narratives of 37 Finnish women and men (in all 80 interviews) and their entire configurations of those specific people who they cite as ‘close or otherwise important’. The analysis stresses the subjective experiences, while also investigating the actualized relational processes and configurations of all personal relationships with certain relationship histories embedded in micro-level structures. The research is based on four empirical sub-studies of personal relationships and a summary discussing the questions of the self and social bond. Discussion draws from G. H. Mead, C. Cooley, N. Elias, T. Scheff, G. Simmel and the contributors of ‘relational sociology’. Sub-studies analyse bonds to others from the viewpoint of biographical disruption and re-configuration of significant others, estranged family bonds, peer support and the formation of the most intimate relationships into exclusive and inclusive configurations. All analyses examine the dialectics of the social and the personal, asking how different structuring mechanisms and personal experiences and negotiations together contribute to the unfolding of the bonds. The summary elaborates personal relationships as social bonds embedded in wider webs of interdependent people and social settings that are laden with cultural expectations. Regarding the question of the relational self, the study proposes both bonding and individuality as significant. They are seen as interdependent phases of the relationality of the self. Bonding anchors the self to its significant relationships, in which individuality is manifested, for example, in contrasting and differentiating dynamics, but also in active attempts to connect with others. Individuality is not a fixed quality of the self, but a fluid and interdependent phase of the relational self. More specifically, it appears in three formats in the flux of relational processes: as a sense of unique self (via cultivation of subjective experiences), as agency and as (a search for) relative autonomy. The study includes an epilogue addressing the ambivalence between the social expectation of individuality in society and the bonded reality of selves.

"Onko yhteisöllisyydellä väliä?" : identiteettiprosessi ja sosiaalinen pääoma epävarmuuden sietämisen merkityksellisinä elementteinä

”Does the community really count? – identity process and social capital as elements in surviving in insecurity and uncertainty” is a combination of five articles. The aim of this study is to answer the question: how or in which ways is it possible to find the role of identity process and social capital in surviving in insecurity and uncertainty? In the introduction part the concepts of community and social capital are examined. Then I will study the articles and try to find out what kinds of elements of identity process and social capital in them can be found in order to survive in the societal change. The study consists of the introduction part and the articles. The articles are: 1. “Is Becoming a Researcher Some Kind of Role-playing” - Roles of the Researcher in the Process of Forming the Identity 2. What Composes Collective Identity in the Polytechnic Community? 3. Opportunities to Succeed or Fear of Failure? -Entrepreneurship from the Youngsters` Point of View 4. Learning Risk-taking Competences 5. “Bricolage”, or Just Putting Things Together? The starting point for the study is the feeling of insecurity that surrounds a person living in the present society: you cannot be sure with whom you are going to co-operate tomorrow. In the “Good Old Days” the harmonious communities “protected” their members and worked strongly toward common aims. Nowadays, partly because of urbanisation, we are so busy that we only have time to take care of ourselves, or rather to say: just of myself. As Bauman (2001) puts it: people turn to communities in which they feel like home. They still long for communality. For Mead (1962) the group and the communality plays a big role: a person needs others to become the whole ”Self.” In acting with others a person can gain much more than working alone (Field 2003). But, as Day (2006) puts it, the reality of community as discovered by empirical reserach is a great deal messier than the abstract and idealized versions used by theorists. Keywords: uncertainty, insecurity, communality, identity process, social capital, significant groups, survival.

From Habits to Social Structures : Pragmatism and Contemporary Social Theory

Pragmatism has sometimes been taken as a catchphrase for epistemological stances in which anything goes. However, other authors argue that the real novelty and contribution of this tradition has to do with its view of action as the context in which all things human take place. Thus, it is action rather than, for example, discourses that should be our starting point in social theory. The introductory section of the book situates pragmatism (especially the ideas of G. H. Mead and John Dewey) within the field and tradition of social theory. This introductory also contextualizes the main core of the book which consists of four chapters. Two of these chapters have been published as articles in scientific journals and one in an edited book. All of them discuss the core problem of social theory: how is action related to social structures (and vice versa)? The argument is that habitual action is the explanation for the emergence of social structures from our action. Action produces structures and social reproduction takes place when action is habitualized; that is, when we develop social dispositions to act in a certain manner in familiar environments. This also means that even though the physical environment is the same for all of us, our habits structure it into different kinds of action possibilities. Each chapter highlights these general insights from different angles. Practice theory has gained momentum in recent years and it has many commonalities with pragmatism because both highlight the situated and corporeal character of human activity. One famous proponent of practice theory is Margaret Archer who has argued that the pragmatism of G. H. Mead leads to an oversocialized conception of selfhood. Mead does indeed present a socialized view of selfhood but this is a meta-sociological argument rather than a substantial sociological claim. Accordingly, one can argue that in this general sense intersubjectivity precedes subjectivity and not the other way around. Such a view does not indicate that our social relation would necessarily "colonize" individual action because there is a place for internal conversations (in Archer s terminology); it is especially in those phases of action where it meets obstacles due to the changes of the environment. The second issue discussed has the background assumption that social structures can fruitfully be conceptualized as institutions. A general classification of different institution theories is presented and it is argued that there is a need for a habitual theory of institutions due to the problems associated with these other theories. So-called habitual institutionalism accounts for institutions in terms of established and prevalent social dispositions that structure our social interactions. The germs of this institution theory can be found in the work of Thorstein Veblen. Since Veblen s times, these ideas have been discussed for example, by the economist Geoffrey M. Hodgson. His ideas on the evolution of institutions are presented but a critical stance is taken towards his tendency of defining institutions with the help of rules because rules are not always present in institutions. Accordingly, habitual action is the most basic but by no means the only aspect of institutional reproduction. The third chapter deals with theme of action and structures in the context of Pierre Bourdieu s thought. Bourdieu s term habitus refers to a system of dispositions which structure social fields. It is argued that habits come close to the concept of habitus in the sense that the latter consists of particular kinds of habits; those that are related to the reproduction of socioeconomic positions. Habits are thus constituents of a general theory of societal reproduction whereas habitus is a systematic combination of socioeconomic habits. The fourth theme relates to issues of social change and development. The capabilities approach has been associated with the name of Amartya Sen, for example, and it underscores problems inhering in economistic ways of evaluating social development. However, Sen s argument has some theoretical problems. For example, his theory cannot adequately confront the problem of relativism. In addition, Sen s discussion lacks also a theory of the role of the public. With the help of arguments derived from pragmatism, one gets an action-based, socially constituted view of freedom in which the role of the public is essential. In general, it is argued that a socially constituted view of agency does not necessarily to lead to pessimistic conclusions about the freedom of action.

Hoivakodin työntekijöiden työhyvinvointi symbolisen interaktionismin näkökulmasta - Työssä kohdattujen ongelmien ja sosiaalisten suhteiden tarkastelua

Tutkielma keskittyy hoivahenkilöstön työhyvinvoinnin merkityksiin ja sen ongelmakohtiin erään vanhusten hoivalaitoksen henkilökunnan kertomana. Tutkimuksen teoreettisena viitekehyksenä toimii Blumerin (1969) teoria symbolisesta interaktionismista, sekä Strykerin strukturaalinen symbolinen interaktionismi (2008). Tutkimuskohteena ovat sosiaaliset prosessit siltä osin kuin ne haittaavat tai tukevat työhyvinvointia hoivalaitoksissa. Tutkielman tavoitteena ei ole käyttää grounded theorya uuden teorian löytämiseen, vaan uusien työhyvinvoinnin tekijöiden kartoittamiseen. Tutkimuskysymykset käsittelivät sitä, millaisena työhyvinvointi näyttäytyy haastateltavien kertomana. Ja lisäksi tarkasteltiin sitä, miten sosiaaliset suhteet työyhteisössä kytkeytyvät työhyvinvoinnin eri aspekteihin. Vanhustenhoidon erityispiirteitä ja aikaisempia työhyvinvoinnin tutkimuksia käydään läpi, sekä kartoitetaan vanhustenhoidon tilaa suomalaisessa yhteiskunnassa. Työhyvinvoinnin käsitettä tarkastellaan sekä yleisesti että vanhustenhoidon osalta. Tutkimusmenetelmänä toimi laadullinen aineistolähtöinen symboliseen interaktionismiin perustuva grounded theory. Aineisto koostui puolistrukturoiduista haastatteluista ja käsitti yhteensä 24 haastattelua, joista 12 valittiin tähän tutkielmaan. Kaikki haastateltavat olivat naisia, johon kuului 8 lähi- ja perushoitajaa, 2 sairaanhoitajaa ja 2 osastonhoitajaa. Haastateltavien valinnan kriteerinä oli työskentely pitkäaikaispotilaiden osastolla. Aineisto analysoitiin glaserilaista grounded theory metodologiaa käyttäen hyödyntäen abduktion logiikkaa. Haastattelut analysoitiin grounded theory metodologian mukaisesti avoimen ja selektiivisen koodauksen avulla CAT-ohjelmaa apuna käyttäen. Tutkimustulosten perusteella havaittiin, että työhyvinvointi koostuu dynaamisista työyhteisön prosesseista, joita säätelevät luottamuksen ja vallankäytön tasapainottelu. Työhyvinvointia kannatteleviin tekijöihin lukeutui avoin kommunikaatio, kollegiaalinen tuki, tasavertaisuus ja yhteiset pelisäännöt. Työhyvinvointia haittaaviksi sosiaalisiksi prosesseiksi fokusoitui vallankäyttö, joka oli selkein työhyvinvointia uhkaava kategoria haastateltavien puheessa. Se oli yhteydessä lukuisiin suoriin ja epäsuoriin interaktion muotoihin, kuten vaientamiseen, syyllistämiseen ja nonkommunikaatioon. Hoivaorganisaation keskeinen dynaaminen elementti oli hoivaorganisaation jähmeys, joka näkyi monella tapaa työyhteisön arjessa vaikeuttaen koko organisaation toimintaa. Organisaation johdolla on merkitystä hoivaorganisaation toiminnan kannalta sekä puheen kulttuurin että konservatiivisen johtamisen muodossa. Merkityssisältöjen erilaisuus johdon ja alaisten välillä aiheutti vuorovaikutuksen vaikeutumista. Sosiaaliset suhteet olivat odotetusti tärkeässä asemassa hoivahenkilöstön työn arjessa. Analyysin tulokset tukivat hoivalaitoksien työntekijöiden työhyvinvointitutkimuksen aikaisempaa linjaa, joissa on korostettu sosiaalisten suhteiden, luottamuksen ja vallankäytön merkitystä työhyvinvointiin. Työn tarkoituksena oli tuottaa uusia työhyvinvoinnin tekijöitä. Tällaisena voidaan pitää nonkommunikaation merkitystä organisaation vuorovaikutusprosessien kannalta. Tutkielman keskeisimmät lähteet ovat: Blumer, H.: Symbolic Interactionism: Perspective and Method; Utriainen, K.: Arvostava vastavuoroisuus ikääntyvien sairaanhoitajien työhyvinvoinnin ytimenä hoitotyössä, sekä Stryker, S.: From Mead to a Structural Symbolic Interactionism and Beyond.

Efficient gradient-free simplex method for estimation of optical properties in image-guided diffuse optical tomography

Typical image-guided diffuse optical tomographic image reconstruction procedures involve reduction of the number of optical parameters to be reconstructed equal to the number of distinct regions identified in the structural information provided by the traditional imaging modality. This makes the image reconstruction problem less ill-posed compared to traditional underdetermined cases. Still, the methods that are deployed in this case are same as those used for traditional diffuse optical image reconstruction, which involves a regularization term as well as computation of the Jacobian. A gradient-free Nelder-Mead simplex method is proposed here to perform the image reconstruction procedure and is shown to provide solutions that closely match ones obtained using established methods, even in highly noisy data. The proposed method also has the distinct advantage of being more efficient owing to being regularization free, involving only repeated forward calculations. (C) 2013 Society of Photo-Optical Instrumentation Engineers (SPIE)

Observations on the spawning of four species on tuna (Neothunnus macropterus, Katsuwonus pelamis, Auxis thazard and Euthynnus lineatus) in the Eastern Pacific Ocean, based on the distribution of their larvae and juveniles

ENGLISH: Knowledge of spawning habits is useful in the elucidation of the life history, ecology and population structure of tropical tunas, and is essential to the sound management of these resources. Until recently, little was known concerning the spawning of tunas, or about the distribution of their larval and juvenile stages, in the Eastern Pacific Ocean. Nichols and Murphy (1944) reported the capture off Colombia of young scombroids ultimately identified as frigate mackerel, Auxis thazard (Schaefer and Marr, 1948a). Fowler (1944) reported the capture off Manzanillo, Mexico of two young tunas, one of which is definitely and the other most likely Neothunnus macropterus (Klawe, 1959). In 1947, young of N. macropterus, K. pelamis, A. thazard and E. lineatus were caught offshore from Central America (Schaefer and Marr, 1948a, 1948b, and Schaefer, 1948). Further collections of young N. macropterus, A. thazard and E. lineatus were made in the same general area in the spring of 1949 (Mead, 1951). In January and February 1955, Clemens (1956) carried Out experiments in rearing young tunas, E. lineatus and A. thazard, in shipboard aquaria, using fish caught off Central America. Matsumoto (1958) reported captures of larval N. macropterus and K. pelamis in the area along the 120th meridian of west longitude. Klawe (1958 and 1961b) reported captures of larval N. macropterus and Auxis from the Revillagigedo Islands. Captures of young Auxis and E. lineatus in the Gulf of Panama in January 1922 during the Dana Expedition have recently been reported by Matsumoto (1959). Capture of juveniles of K. pelamis, E. lineatus and Auxis in the area off tropical Mexico and in the area of outlying islands during the SCOT Expedition has been reported by Klawe (1960a). SPANISH: El conocimiento sobre los hábitos del desove es útil para el esclarecimiento de la historia natural, ecología y estructura de las poblaciones de atunes tropicales, y es esencial para la acertada administración de estos recursos. Hasta hace poco tiempo no se sabía mucho sobre el desove de los atunes o acerca de la distribución de sus larvas y juveniles en el Océano Pacífico Oriental. Nichols y Murphy (1944) informaron sobre la captura frente a Colombia de escómbridos jóvenes últimamente identificados como melva, Auxis thazard (Schaefer y Marr, 1948a). Fowler (1944) también informó sobre la captura de dos atunes jóvenes frente a Manzanillo, México, uno de los cuales era definitivamente Neothunnus macropterus y el otro era lo más probable que también lo fuera (Klawe, 1959). En 1947 se capturaron especímenes juveniles de N. macropterus, K. pelamis, A. thazard y E. lineatus frente a la América Central (Schaefer y Marr, 1948a, 1948b, y Schaefer, 1948). Otras recolecciones de ejemplares jóvenes de N. macropterus, A. thazard y E. lineatus fueron hechas en la misma área general durante la primavera de 1949 (Mead, 1951). En enero y febrero de 1955, Clemens (1956) efectuó experimentos de crianza de atunes jóvenes, E. lineatus y A.. thazard, en acuarios a bordo para lo que empleó peces capturados frente a la América Central. Matsumoto (1958) informó sobre capturas de larvas de N. macropterus y K. pelamis en el área a lo largo del meridiano 120 de longitud oeste. Klawe (1958 y 1961b) ha dado cuenta también de capturas de larvas de N. macropterus y Auxis en las Islas Revillagigedo. Matsumoto (1959) ha informado recientemente acerca de capturas de ejemplares jóvenes de Auxis y E. lineatus en el Golfo de Panamá en enero de 1922 durante la Expedición Dana. Klawe (1960a) informó así mismo que durante la Expedición SCOT se capturaron juveniles de K. pelamis, E. lineatus y Auxis en el área frente a la zona tropical de México y en la región de las islas alejadas del continente.

Triangle Evolution-A Hybrid Heuristic for Global Optimization

Abstract This paper presents a hybrid heuristic{triangle evolution (TE) for global optimization. It is a real coded evolutionary algorithm. As in di®erential evolution (DE), TE targets each individual in current population and attempts to replace it by a new better individual. However, the way of generating new individuals is di®erent. TE generates new individuals in a Nelder- Mead way, while the simplices used in TE is 1 or 2 dimensional. The proposed algorithm is very easy to use and e±cient for global optimization problems with continuous variables. Moreover, it requires only one (explicit) control parameter. Numerical results show that the new algorithm is comparable with DE for low dimensional problems but it outperforms DE for high dimensional problems.

基于CFD分析的弧形翼导弹气动外形优化

为探索将高精度CFD分析引入高超声速飞行器气动外形优化设计的可行性,以6°飞行攻角、6马赫飞行速度下的最大升阻比为目标,引入CFD分析计算气动性能,使用Nelder-Mead可变多面体搜索方法对高超声速弧形翼导弹进行了气动外形优化.计算结果表明,在满足弹翼安全性的条件下,优化外形比初始外形的升阻比有明显提高(约9.22%),同时阻力系数、弹翼展长和弹翼面积均有不同程度的降低.计算结果充分证明了优化设计的有效性和采用CFD分析的必要性.依据该文结果提出一种前掠弧形翼高超声速导弹气动布局方案.

Optimal Fuzzy PID Controller with Incomplete Derivation and Its Simulation Research on Application of Intelligent Artificial Legs

提出了一种新的最优模糊PID控制器，它由两部分组成，即在线模糊推理机构和带有不完全微分的常规PID控制器，在模糊推理机构中，引入了三个可调节因子xp,xi和xd，其作用是进一步修改和优化模糊推理的结果，以使控制器对一个给定对象具有最优的控制效果，可调节因子的最优值采用ITAE准则及Nelder和Mead提出的柔性多面体最优搜索算法加以确定，这种PID控制器被用来控制由作者设计的智能人工腿中的一个直流电机，仿真结果表明该控制器的设计是非常有效的，它可被用于控制各种不同的对象和过程。

A Comparison of Hardware Implementations for Low-Level Vision Algorithms

Early and intermediate vision algorithms, such as smoothing and discontinuity detection, are often implemented on general-purpose serial, and more recently, parallel computers. Special-purpose hardware implementations of low-level vision algorithms may be needed to achieve real-time processing. This memo reviews and analyzes some hardware implementations of low-level vision algorithms. Two types of hardware implementations are considered: the digital signal processing chips of Ruetz (and Broderson) and the analog VLSI circuits of Carver Mead. The advantages and disadvantages of these two approaches for producing a general, real-time vision system are considered.

A cognitive approach to identifying measurable milestones for programming skill acquisition

Mead, J., Gray, S., Hamer, J., James, R., Sorva, J., Clair, C. S., and Thomas, L. 2006. A cognitive approach to identifying measurable milestones for programming skill acquisition. SIGCSE Bull. 38, 4 (Dec. 2006), 182-194.

The Ontogeny of Mucosal and Systemic Antibody Responses to HIV-1 Infection

The humoral immune system plays a critical role in the clearance of numerous pathogens. In the setting of HIV-1 infection, the virus infects, integrates its genome into the host's cells, replicates, and establishes a reservoir of virus-infected cells. The initial antibody response to HIV-1 infection is targeted to non-neutralizing epitopes on HIV-1 Env gp41, and when a neutralizing response does develop months after transmission, it is specific for the autologous founder virus and the virus escapes rapidly. After continuous waves of antibody mediated neutralization and viral escape, a small subset of infected individuals eventually develop broad and potent heterologous neutralizing antibodies years after infection. In this dissertation, I have studied the ontogeny of mucosal and systemic antibody responses to HIV-1 infection by means of three distinct aims: 1. Determine the origin of the initial antibody response to HIV-1 infection. 2. Characterize the role of restricted VH and VL gene segment usage in shaping the antibody response to HIV-1 infection. 3. Determine the role of persistence of B cell clonal lineages in shaping the mutation frequencies of HIV-1 reactive antibodies.

After the introduction (Chapter 1) and methods (Chapter 2), Chapter 3 of this dissertation describes a study of the antibody response of terminal ileum B cells to HIV-1 envelope (Env) in early and chronic HIV-1 infection and provides evidence for the role of environmental antigens in shaping the repertoire of B cells that respond to HIV-1 infection. Previous work by Liao et al. demonstrated that the initial plasma cell response in the blood to acute HIV-1 infection is to gp41 and is derived from a polyreactive memory B cell pool. Many of these antibodies cross-reacted with commensal bacteria, Therefore, in Chapter 3, the relationship of intestinal B cell reactivity with commensal bacteria to HIV-1 infection-induced antibody response was probed using single B cell sorting, reverse transcription and nested polymerase chain reaction (RT- PCR) methods, and recombinant antibody technology. The dominant B cell response in the terminal ileum was to HIV-1 envelope (Env) gp41, and 82% of gp41- reactive antibodies cross-reacted with commensal bacteria whole cell lysates. Pyrosequencing of blood B cells revealed HIV-1 antibody clonal lineages shared between ileum and blood. Mutated IgG antibodies cross-reactive with both Env gp41 and commensal bacteria could also be isolated from the terminal ileum of HIV-1 uninfected individuals. Thus, the antibody response to HIV-1 can be shaped by intestinal B cells stimulated by commensal bacteria prior to HIV-1 infection to develop a pre-infection pool of memory B cells cross-reactive with HIV-1 gp41.

Chapter 4 details the study of restricted VH and VL gene segment usage for gp41 and gp120 antibody induction following acute HIV-1 infection; mutations in gp41 lead to virus enhanced neutralization sensitivity. The B cell repertoire of antibodies induced in a HIV-1 infected African individual, CAP206, who developed broadly neutralizing antibodies (bnAbs) directed to the HIV-1 envelope gp41 membrane proximal external region (MPER), is characterized. Understanding the selection of virus mutants by neutralizing antibodies is critical to understanding the role of antibodies in control of HIV-1 replication and prevention from HIV-1 infection. Previously, an MPER neutralizing antibody, CAP206-CH12, with the binding footprint identical to that of MPER broadly neutralizing antibody 4E10, that like 4E10 utilized the VH1-69 and VK3-20 variable gene segments was isolated from this individual (Morris et al., 2011). Using single B cell sorting, RT- PCR methods, and recombinant antibody technology, Chapter 4 describes the isolation of a VH1-69, Vk3-20 glycan-dependent clonal lineage from CAP206, targeted to gp120, that has the property of neutralizing a neutralization sensitive CAP206 transmitted/founder (T/F) and heterologous viruses with mutations at amino acids 680 or 681 in the MPER 4E10/CH12 binding site. These data demonstrate sites within the MPER bnAb epitope (aa 680-681) in which mutations can be selected that lead to viruses with enhanced sensitivity to autologous and heterologous neutralizing antibodies.

In Chapter 5, I have completed a comparison of evolution of B cell clonal lineages in two HIV-1 infected individuals who have a predominant VH1-69 response to HIV-1 infection--one who produces broadly neutralizing MPER-reactive mAbs and one who does not. Autologous neutralization in the plasma takes ~12 weeks to develop (Gray et al., 2007; Tomaras et al., 2008b). Only a small subset of HIV-1 infected individuals develops high plasma levels of broad and potent heterologous neutralization, and when it does occur, it typically takes 3-4 years to develop (Euler et al., 2010; Gray et al., 2007; 2011; Tomaras et al., 2011). The HIV-1 bnAbs that have been isolated to date have a number of unusual characteristics including, autoreactivity and high levels of somatic hypermutations, which are typically tightly regulated by immune control mechanisms (Haynes et al., 2005; 2012b; Kwong and Mascola, 2012; Scheid et al., 2009a). The VH mutation frequencies of bnAbs average ~15% but have been shown to be as high as 32% (reviewed in Mascola and Haynes, 2013; Kwong and Mascola, 2012). The high frequency of somatic hypermutations suggests that the B cell clonal lineages that eventually produce bnAbs undergo high-levels of affinity maturation, implying prolonged germinal center (GC) reactions and high levels of T cell help. To study the duration of HIV-1- reactive B cell clonal persistence, HIV-1 reactive and non HIV-1- reactive B cell clonal lineages were isolated from an HIV-1 infected individual that produces bnAbs, CAP206, and an HIV-1 infected individual who does not produce bnAbs, 004-0. Single B cell sorting, RT-PCR and recombinant antibody technology was used to isolate and produce monoclonal antibodies from multiple time points from each individual. B cell sequences clonally related to mAbs isolated by single cell PCR were identified within pyrosequences of longitudinal samples of these two individuals. Both individuals produced long-lived B cell clones that persisted from 0-232 weeks in CAP206, and 0-238 weeks in 004-0. The average length of persistence of clones containing members isolated from two separate time points was 91.5 weeks both individuals. Examples of the continued evolution of clonal lineages were observed in both the bnAb and non-bnAb individual. These data indicated that the ability to generate persistent and evolving B cell clonal lineages occurs in both bnAb and non-bnAb individuals, suggesting that some alternative host or viral factor is critical for the generation of highly mutated broadly neutralizing antibodies.

Together the studies described in Chapter 3-5 show that multiple factors influence the antibody response to HIV-1 infection. The initial antibody response to HIV-1 Env gp41 can be shaped by a B cell response to intestinal commensal bacteria prior to HIV-1 infection. VH and VL gene segment restriction can impact the B cell response to multiple HIV-1 antigens, and virus escape mutations in the MPER can confer enhanced neutralization sensitivity to autologous and heterologous antibodies. Finally, the ability to generate long-lived HIV-1 clonal lineages in and of itself does not confer on the host the ability to produce bnAbs.