Concomitantly Diagnosed Disseminated M kansasii Infection and Hairy Cell Leukemia With Review of Pathophysiology.

The association between Hairy Cell Leukemia (HCL) and non-tuberculous mycobacterial infections (NTMs) is well described, most notably Mycobacterium kansasii. The exact pathophysiology is not known. We report a case of a 31-year-old male with concomitantly diagnosed HCL and disseminated M kansasii infection who presented with rash, pancytopenia, and bulky axillary lymphadenopathy. The M kansasii was initially diagnosed through use of cell-free DNA detection and confirmed by bone marrow and lymph node cultures. Hairy Cell Leukemia was diagnosed with peripheral flow cytometry and confirmed via the same bone marrow sample. His HCL was put into remission with a single course of cladribine and rituximab chemotherapy; however, his M kansasii infection persisted for 6 months despite aggressive antimicrobial and surgical therapy. It was finally controlled using high-dose rifampin in combination with azithromycin and ethambutol. This case highlights the known link between HCL and M kansasii. Furthermore, it hints at potential causes beyond chemotherapy-induced immunocompromise. Notable possibilities include HCL cells acting as sanctuary sites for M kansasii to evade the immune system, and subclinical M kansasii infections causing NLRP3 inflammasome overactivation to trigger the oncogenic transformation to HCL. More research into the pathophysiologic link between HCL and M kansasii infections would allow for more effective prevention, diagnosis, and treatment of these severe atypical infections which are the major cause of morbidity in the cladribine era of HCL treatment.

Impact of changing climate on bryophyte contributions to terrestrial water, carbon, and nitrogen cycles.

Bryophytes, including the lineages of mosses, liverworts, and hornworts, are the second-largest photoautotroph group on Earth. Recent work across terrestrial ecosystems has highlighted how bryophytes retain and control water, fix substantial amounts of carbon (C), and contribute to nitrogen (N) cycles in forests (boreal, temperate, and tropical), tundra, peatlands, grasslands, and deserts. Understanding how changing climate affects bryophyte contributions to global cycles in different ecosystems is of primary importance. However, because of their small physical size, bryophytes have been largely ignored in research on water, C, and N cycles at global scales. Here, we review the literature on how bryophytes influence global biogeochemical cycles, and we highlight that while some aspects of global change represent critical tipping points for survival, bryophytes may also buffer many ecosystems from change due to their capacity for water, C, and N uptake and storage. However, as the thresholds of resistance of bryophytes to temperature and precipitation regime changes are mostly unknown, it is challenging to predict how long this buffering capacity will remain functional. Furthermore, as ecosystems shift their global distribution in response to changing climate, the size of different bryophyte-influenced biomes will change, resulting in shifts in the magnitude of bryophyte impacts on global ecosystem functions.

Thallus hydrophobicity: A low-cost method for understanding lichen ecophysiological responses to environmental changes.

Premise: Methods to evaluate lichen thalli hydrophobicity have previously been described, but only recently has hydrophobicity been shown to be an important functional trait related to water regulation dynamics that could be used to predict future climate change effects. We describe a novel protocol to measure lichen thallus hydrophobicity that aims to be an easier and more affordable approach. Methods and Results: Our protocol requires only a micropipette, distilled water, a tripod, and a smartphone or camera. Hydrophobicity is inferred from multiple metrics associated with the absorption times of standardized droplets (initial and total absorption time). We used a data set of 93 lichen taxa with different growth forms and from different biomes and demonstrated that this method is well suited for capturing different levels of hydrophobicity, including very hydrophilic species. Conclusions: Our results show that this new protocol to measure lichen hydrophobicity is a rapid and low-cost method to assess an ecophysiologically based functional trait that can be used with almost no limitations, including in different climates, lichen species, and growth forms.

Premisa: En el pasado se han descrito métodos para evaluar la hidrofobicidad de los talos liquénicos, sin embargo, no fue hasta recientemente que se demostró la importancia de esta propiedad como rasgo funcional en relación a la dinámica de regulación hídrica de los talos, permitiendo utilizarla como herramienta para predecir futuros efectos del cambio climático. Describimos un nuevo protocolo para medir la hidrofobicidad de los talos liquénicos que pretende ser más fácil y asequible. Métodos y Resultados: Nuestro protocolo requiere solamente de una micropipeta, agua destilada, un trípode y un teléfono o una cámara. La hidrofobicidad es inferida a partir de múltiples métricas asociadas a los tiempos de absorción de gotas de un volumen estandarizado (tiempo de absorción inicial y tiempo de absorción total). Utilizamos datos de 93 taxones de líquenes con diferentes formas de crecimiento y provenientes de distintos biomas, lo que permitió demostrar que este método es adecuado para capturar diferentes niveles de hidrofobicidad, incluyendo a especies altamente hidrofílicas. Conclusiones: Nuestros resultados muestran que la medición de la hidrofobicidad de los talos es un método rápido y de bajo costo que permite evaluar un rasgo funcional basado en la ecofisiología de líquenes y que puede ser utilizado prácticamente sin limitaciones, incluyendo en diferentes climas, especies y formas de crecimiento.

Moss functional trait ecology: Trends, gaps, and biases in the current literature.

Functional traits are critical tools in plant ecology for capturing organism-environment interactions based on trade-offs and making links between organismal and ecosystem processes. While broad frameworks for functional traits have been developed for vascular plants, we lack the same for bryophytes, despite an escalation in the number of studies on bryophyte functional trait in the last 45 years and an increased recognition of the ecological roles bryophytes play across ecosystems. In this review, we compiled data from 282 published articles (10,005 records) that focused on functional traits measured in mosses and sought to examine trends in types of traits measured, capture taxonomic and geographic breadth of trait coverage, reveal biases in coverage in the current literature, and develop a bryophyte-function index (BFI) to describe the completeness of current trait coverage and identify global gaps to focus research efforts. The most commonly measured response traits (those related to growth/reproduction in individual organisms) and effect traits (those that directly affect community/ecosystem scale processes) fell into the categories of morphology (e.g., leaf area, shoot height) and nutrient storage/cycling, and our BFI revealed that these data were most commonly collected from temperate and boreal regions of Europe, North America, and East Asia. However, fewer than 10% of known moss species have available functional trait information. Our synthesis revealed a need for research on traits related to ontogeny, sex, and intraspecific plasticity and on co-measurement of traits related to water relations and bryophyte-mediated soil processes.

A new species of the fish louse genus Dipteropeltis Calman, 1912 (Crustacea: Branchiura) from Peru.

Dipteropeltis is a poorly described genus of fish louse endemic to South America. In a small blackwater region within Loreto, Peru, 13 adult and juvenile specimens of an unidentified species of Dipteropeltis Calman, 1912, as well as one adult specimen of D. hirundo Calman, 1912, were observed and collected. Scanning electron and light micrographs were acquired to examine and measure key features of these specimens. Morphological differences from the two known species of Dipteropeltis, D. hirundo and D. campanaformis Neethling et al., 2014, indicate that the collected specimens represent a new species. Dipteropeltis longicaudatus sp. nov. is diagnosed by elongate abdominal lobes, a chevron-shaped carapace, and uniquely shaped maxillae. One specimen represents the longest branchiuran documented to date at 31.5 mm. Additionally, we provide the first sequence data for this genus using DNA barcoding, which corroborates our designation of a new species. Videos were also captured that document behaviors including host attachment, pulsating abdominal lobes, suction disc "walking", and swimming. Findings have implications for its teleost hosts, Triportheus albus Cope, 1872 and Brycon amazonicus Spix & Agassiz, 1829, the latter being a critical species for aquaculture and commercial fisheries in Amazonia.

Absence of a molecular circadian clock in the preimplantation embryo is a conserved characteristic in the mammal.

In brief: It is not known when a functional circadian clock is established in the developing embryo. Lack of expression of key genes involved in the clock mechanism is indicative that a functional circadian clock mechanism is absent in the mammalian preimplantation embryo through the blastocyst stage of development. Abstract: An embryonic circadian clock could conceivably organize cellular and developmental events temporally and in synchrony with other circadian rhythms in the mother. The hypothesis that a functional molecular clock exists in the preimplantation bovine, pig, human, and mouse embryo was tested by using publicly available RNAseq datasets to examine developmental changes in expression of the core genes responsible for the circadian clock - CLOCK, ARNTL, PER1, PER2, CRY1, and CRY2. In general, the transcript abundance of each gene decreased as development advanced to the blastocyst stage. The most notable exception was for CRY2, where transcript abundance was low and constant from the two-cell or four-cell to the blastocyst stage. Developmental patterns were generally the same for all species although there were some species-specific patterns such as an absence of PER1 expression in the pig, an increase in ARNTL expression at the four-cell stage in human, and an increase in expression of Clock and Per1 from the zygote to two-cell stage in the mouse. Analysis of intronic reads (indicative of embryonic transcription) for bovine embryos indicated an absence of embryonic transcription. Immunoreactive CRY1 was not detected in the bovine blastocyst. Results indicate that the preimplantation mammalian embryo lacks a functional intrinsic clock although specific components of the clock mechanism could conceivably play a role in other functions in the embryo.

Lichen ecophysiology in a changing climate.

Lichens are one of the most iconic and ubiquitous symbioses known, widely valued as indicators of environmental quality and, more recently, climate change. Our understanding of lichen responses to climate has greatly expanded in recent decades, but some biases and constraints have shaped our present knowledge. In this review we focus on lichen ecophysiology as a key to predicting responses to present and future climates, highlighting recent advances and remaining challenges. Lichen ecophysiology is best understood through complementary whole-thallus and within-thallus scales. Water content and form (vapor or liquid) are central to whole-thallus perspectives, making vapor pressure differential (VPD) a particularly informative environmental driver. Responses to water content are further modulated by photobiont physiology and whole-thallus phenotype, providing clear links to a functional trait framework. However, this thallus-level perspective is incomplete without also considering within-thallus dynamics, such as changing proportions or even identities of symbionts in response to climate, nutrients, and other stressors. These changes provide pathways for acclimation, but their understanding is currently limited by large gaps in our understanding of carbon allocation and symbiont turnover in lichens. Lastly, the study of lichen physiology has mainly prioritized larger lichens at high latitudes, producing valuable insights but underrepresenting the range of lichenized lineages and ecologies. Key areas for future work include improving geographic and phylogenetic coverage, greater emphasis on VPD as a climatic factor, advances in the study of carbon allocation and symbiont turnover, and the incorporation of physiological theory and functional traits in our predictive models.

Carbon-concentrating mechanisms are a key trait in lichen ecology and distribution.

Carbon-concentrating mechanisms (CCMs) are a widespread phenomenon in photosynthetic organisms. In vascular plants, the evolution of CCMs ([C44-carbon compound] and crassulacean acid metabolism [CAM]) is associated with significant shifts, most often to hot, dry and bright, or aquatic environments. If and how CCMs drive distributions of other terrestrial photosynthetic organisms, remains little studied. Lichens are ecologically important obligate symbioses between fungi and photosynthetic organisms. The primary photosynthetic partner in these symbioses can include CCM-presenting cyanobacteria (as carboxysomes), CCM-presenting green algae (as pyrenoids) or green algae lacking any CCM. We use an extensive dataset of lichen communities from eastern North America, spanning a wide climatic range, to test the importance of CCMs as predictors of lichen ecology and distribution. We show that the presence or absence of CCMs leads to opposite responses to temperature and precipitation in green algal lichens, and different responses in cyanobacterial lichens. These responses contrast with our understanding of lichen physiology, whereby CCMs mitigate carbon limitation by water saturation at the cost of efficient use of vapor hydration. This study demonstrates that CCM status is a key functional trait in obligate lichen symbioses, equivalent in importance to its role in vascular plants, and central for studying present and future climate responses.

Climate warming causes photobiont degradation and carbon starvation in a boreal climate sentinel lichen.

PREMISE: The long-term potential for acclimation by lichens to changing climates is poorly known, despite their prominent roles in forested ecosystems. Although often considered "extremophiles," lichens may not readily acclimate to novel climates well beyond historical norms. In a previous study (Smith et al., 2018), Evernia mesomorpha transplants in a whole-ecosystem climate change experiment showed drastic mass loss after 1 yr of warming and drying; however, the causes of this mass loss were not addressed. METHODS: We examined the causes of this warming-induced mass loss by measuring physiological, functional, and reproductive attributes of lichen transplants. RESULTS: Severe loss of mass and physiological function occurred above +2°C of experimental warming. Loss of algal symbionts ("bleaching") and turnover in algal community compositions increased with temperature and were the clearest impacts of experimental warming. Enhanced CO2 had no significant physiological or symbiont composition effects. The functional loss of algal photobionts led to significant loss of mass and specific thallus mass (STM), which in turn reduced water-holding capacity (WHC). Although algal genotypes remained detectable in thalli exposed to higher stress, within-thallus photobiont communities shifted in composition toward greater diversity. CONCLUSIONS: The strong negative impacts of warming and/or lower humidity on Evernia mesomorpha were driven by a loss of photobiont activity. Analogous to the effects of climate change on corals, the balance of symbiont carbon metabolism in lichens is central to their resilience to changing conditions.

Insights into the mechanism of Huanglongbing tolerance in the Australian finger lime (Citrus australasica).

The Australian finger lime (Citrus australasica) is tolerant to Huanglongbing (HLB; Citrus greening). This species can be utilized to develop HLB tolerant citrus cultivars through conventional breeding and biotechnological approaches. In this report, we conducted a comprehensive analysis of transcriptomic data following a non-choice infection assay to understand the CaLas tolerance mechanisms in the finger lime. After filtering 3,768 differentially expressed genes (DEGs), 2,396 were downregulated and 1,372 were upregulated in CaLas-infected finger lime compared to CaLas-infected HLB-susceptible 'Valencia' sweet orange. Comparative analyses revealed several DEGs belonging to cell wall, ß-glucanase, proteolysis, R genes, signaling, redox state, peroxidases, glutathione-S-transferase, secondary metabolites, and pathogenesis-related (PR) proteins categories. Our results indicate that the finger lime has evolved specific redox control systems to mitigate the reactive oxygen species and modulate the plant defense response. We also identified candidate genes responsible for the production of Cys-rich secretory proteins and Pathogenesis-related 1 (PR1-like) proteins that are highly upregulated in infected finger lime relative to noninfected and infected 'Valencia' sweet orange. Additionally, the anatomical analysis of phloem and stem tissues in finger lime and 'Valencia' suggested better regeneration of phloem tissues in finger lime in response to HLB infection. Analysis of callose formation following infection revealed a significant difference in the production of callose plugs between the stem phloem of CaLas+ 'Valencia' sweet orange and finger lime. Understanding the mechanism of resistance will help the scientific community design strategies to protect trees from CaLas infection and assist citrus breeders in developing durable HLB tolerant citrus varieties.

Proteomic and bioinformatic analyses of proteins in the outer membrane and extracellular compartments and outer membrane vesicles of Candidatus Liberibacter species.

Citrus Huanglongbing (HLB) is the most devastating citrus disease in the world. Candidatus Liberibacter asiaticus (Las) is the prevalent HLB pathogen, which is yet to be cultivated. A recent study demonstrates that Las does not contain pathogenicity factors that are directly responsible for HLB symptoms. Instead, Las triggers systemic and chronic immune responses, representing a pathogen-triggered immune disease. Importantly, overproduction of reactive oxygen species (ROS) causes systemic cell death of phloem tissues, thus causing HLB symptoms. Because Las resides in the phloem tissues, it is expected that phloem cell might recognize outer membrane proteins, outer membrane vesicle (OMV) proteins and extracellular proteins of Las to contribute to the immune responses. Because Las has not been cultivated, we used Liberibacter crescens (Lcr) as a surrogate to identify proteins in the OM fraction, OMV proteins and extracellular proteins by liquid chromatography with tandem mass spectrometry (LC-MS/MS). We observed OMVs of Lcr under scanning electron microscope, representing the first experimental evidence that Liberibacter can deliver proteins to the extracellular compartment. In addition, we also further analyzed LC-MS/MS data using bioinformatic tools. Our study provides valuable information regarding the biology of Ca. Liberibacter species and identifies many putative proteins that may interact with host proteins in the phloem tissues.

Step in Time: Conservation of Circadian Clock Genes in Animal Evolution.

Over the past few decades, the molecular mechanisms responsible for circadian phenotypes of animals have been studied in increasing detail in mammals, some insects, and other invertebrates. Particular circadian proteins and their interactions are shared across evolutionary distant animals, resulting in a hypothesis for the canonical circadian clock of animals. As the number of species for which the circadian clockwork has been described increases, the circadian clock in animals driving cyclical phenotypes becomes less similar. Our focus in this review is to develop and synthesize the current literature to better understand the antiquity and evolution of the animal circadian clockwork. Here, we provide an updated understanding of circadian clock evolution in animals, largely through the lens of conserved genes characterized in the circadian clock identified in bilaterian species. These comparisons reveal extensive variation within the likely composition of the core clock mechanism, including losses of many genes, and that the ancestral clock of animals does not equate to the bilaterian clock. Despite the loss of these core genes, these species retain circadian behaviors and physiology, suggesting novel clocks have evolved repeatedly. Additionally, we highlight highly conserved cellular processes (e.g., cell division, nutrition) that intersect with the circadian clock of some animals. The conservation of these processes throughout the animal tree remains essentially unknown, but understanding their role in the evolution and maintenance of the circadian clock will provide important areas for future study.

Evolutionary biology of lichen symbioses.

Lichens are the symbiotic outcomes of open, interspecies relationships, central to which are a fungus and a phototroph, typically an alga and/or cyanobacterium. The evolutionary processes that led to the global success of lichens are poorly understood. In this review, we explore the goods and services exchange between fungus and phototroph and how this propelled the success of both symbiont and symbiosis. Lichen fungal symbionts count among the only filamentous fungi that expose most of their mycelium to an aerial environment. Phototrophs export carbohydrates to the fungus, which converts them to specific polyols. Experimental evidence suggests that polyols are not only growth and respiratory substrates but also play a role in anhydrobiosis, the capacity to survive desiccation. We propose that this dual functionality is pivotal to the evolution of fungal symbionts, enabling persistence in environments otherwise hostile to fungi while simultaneously imposing costs on growth. Phototrophs, in turn, benefit from fungal protection from herbivory and light stress, while appearing to exert leverage over fungal sex and morphogenesis. Combined with the recently recognized habit of symbionts to occur in multiple symbioses, this creates the conditions for a multiplayer marketplace of rewards and penalties that could drive symbiont selection and lichen diversification.

A cationic lipid mediated CRISPR/Cas9 technique for the production of stable genome edited citrus plants.

BACKGROUND: The genetic engineering of crops has enhanced productivity in the face of climate change and a growing global population by conferring desirable genetic traits, including the enhancement of biotic and abiotic stress tolerance, to improve agriculture. The clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) system has been found to be a promising technology for genomic editing. Protoplasts are often utilized for the development of genetically modified plants through in vitro integration of a recombinant DNA fragment into the plant genome. We targeted the citrus Nonexpressor of Pathogenesis-Related 3 (CsNPR3) gene, a negative regulator of systemic acquired resistance (SAR) that governs the proteasome-mediated degradation of NPR1 and developed a genome editing technique targeting citrus protoplast DNA to produce stable genome-edited citrus plants. RESULTS: Here, we determined the best cationic lipid nanoparticles to deliver donor DNA and described a protocol using Lipofectamine™ LTX Reagent with PLUS Reagent to mediate DNA delivery into citrus protoplasts. A Cas9 construct containing a gRNA targeting the CsNPR3 gene was transfected into citrus protoplasts using the cationic lipid transfection agent Lipofectamine with or without polyethylene glycol (PEG, MW 6000). The optimal transfection efficiency for the encapsulation was 30% in Lipofectamine, 51% in Lipofectamine with PEG, and 2% with PEG only. Additionally, plasmid encapsulation in Lipofectamine resulted in the highest cell viability percentage (45%) compared with PEG. Nine edited plants were obtained and identified based on the T7EI assay and Sanger sequencing. The developed edited lines exhibited downregulation of CsNPR3 expression and upregulation of CsPR1. CONCLUSIONS: Our results demonstrate that utilization of the cationic lipid-based transfection agent Lipofectamine is a viable option for the successful delivery of donor DNA and subsequent successful genome editing in citrus.

Citrus Huanglongbing is a pathogen-triggered immune disease that can be mitigated with antioxidants and gibberellin.

Huanglongbing (HLB) is a devastating disease of citrus, caused by the phloem-colonizing bacterium Candidatus Liberibacter asiaticus (CLas). Here, we present evidence that HLB is an immune-mediated disease. We show that CLas infection of Citrus sinensis stimulates systemic and chronic immune responses in phloem tissue, including callose deposition, production of reactive oxygen species (ROS) such as H2O2, and induction of immunity-related genes. The infection also upregulates genes encoding ROS-producing NADPH oxidases, and downregulates antioxidant enzyme genes, supporting that CLas causes oxidative stress. CLas-triggered ROS production localizes in phloem-enriched bark tissue and is followed by systemic cell death of companion and sieve element cells. Inhibition of ROS levels in CLas-positive stems by NADPH oxidase inhibitor diphenyleneiodonium (DPI) indicates that NADPH oxidases contribute to CLas-triggered ROS production. To investigate potential treatments, we show that addition of the growth hormone gibberellin (known to have immunoregulatory activities) upregulates genes encoding H2O2-scavenging enzymes and downregulates NADPH oxidases. Furthermore, foliar spray of HLB-affected citrus with gibberellin or antioxidants (uric acid, rutin) reduces H2O2 concentrations and cell death in phloem tissues and reduces HLB symptoms. Thus, our results indicate that HLB is an immune-mediated disease that can be mitigated with antioxidants and gibberellin.

Morphometric and structural analysis of Florida manatee spermatozoa.

Sperm characteristics, such as sperm morphology and sperm morphometry are important in assessing sperm quality. This is especially important for the management and conservation of endangered and exotic species, like the Florida manatee, where information of this nature is extremely limited. In this study, we fill this knowledge gap to better understand the reproductive physiology of Florida manatees by conducting the first extensive analysis of sperm morphometry and ultrastructure. Sperm were retrieved from the vas deferens of nine recently deceased Florida manatees. Computer-aided sperm morphology analysis (CASMA) was used for morphometric analysis and laser-scanning confocal microscopy and electron microscopy were used for structural and ultrastructural characterization. Our findings reveal new morphometric and structural data for the Florida manatee spermatozoon. Twelve morphometric features of Florida manatee sperm were quantified with some approximately 1.5-2 times larger than those previously reported. Ultrastructurally, the Florida manatee spermatozoon followed a mammalian structural pattern with an ovate-shaped head, midpiece containing 84-90 mitochondria, and a flagellum. However, unique ultrastructural features were identified. Distinct, rectangular-like enlargement of four outer dense fibers surrounding the axoneme was evident, which may provide additional tensile strength to counteract the forces on sperm transiting the female reproductive tract. Likewise, strong localization of F-actin fibers within the midpiece may function to maintain sperm integrity within the female reproductive tract. These findings highlight the potential effects of sexual selective pressures on sperm size and structure in the Florida manatee and provide avenues for research on the occurrence of sperm competition in this species.

Differential retention contributes to racial/ethnic disparity in U.S. academia.

Several racial and ethnic identities are widely understood to be under-represented within academia, however, actual quantification of this under-representation is surprisingly limited. Challenges include data availability, demographic inertia and identifying comparison points. We use de-aggregated data from the U.S. National Science Foundation to construct a null model of ethnic and racial representation in one of the world's largest academic communities. Making comparisons between our model and actual representation in academia allows us to measure the effects of retention (while controlling for recruitment) at different academic stages. We find that, regardless of recruitment, failed retention contributes to mis-representation across academia and that the stages responsible for the largest disparities differ by race and ethnicity: for Black and Hispanic scholars this occurs at the transition from graduate student to postdoctoral researcher whereas for Native American/Alaskan Native and Native Hawaiian/Pacific Islander scholars this occurs at transitions to and within faculty stages. Even for Asian and Asian-Americans, often perceived as well represented, circumstances are complex and depend on choice of baseline. Our findings demonstrate that while recruitment continues to be important, retention is also a pervasive barrier to proportional representation. Therefore, strategies to reduce mis-representation in academia must address retention. Although our model does not directly suggest specific strategies, our framework could be used to project how representation in academia might change in the long-term under different scenarios.

Utilization of somatic fusion techniques for the development of HLB tolerant breeding resources employing the Australian finger lime (Citrus australasica).

The Australian finger lime is a unique citrus species that has gained importance due to its unique fruit characteristics and perceived tolerance to Huanglongbing (HLB), an often-fatal disease of citrus trees. In this study, we developed allotetraploid finger lime hybrids and cybrids by utilizing somatic cell fusion techniques to fuse diploid 'OLL8' sweet orange or 'Page' tangelo callus-derived protoplasts with finger lime (FL) mesophyll-derived protoplasts. Six somatic fusions were regenerated from the 'OLL8' + FL fusion, while three putative cybrids were regenerated from the 'Page' + FL fusion. Ploidy levels and nuclear-expressed sequence tag derived simple sequence repeat (EST-SSR) markers confirmed the somatic hybrid production, and mitochondrial DNA primer sets confirmed the cybrid nature. Several trees produced by the somatic fusion remained HLB negative even after 6 years of growth in an HLB-endemic environment. Pathogenesis related (PR) and other genes that are often upregulated in HLB-tolerant trees were also upregulated in our somatic fusions. These newly developed somatic fusions and cybrids could potentially be used as breeding parents to develop the next generation of improved HLB-tolerant rootstocks and scions.

Quadrilateral space syndrome induced by a large degenerative osteophyte.

A 41-year-old gentleman presented with decreased range of motion. Initial radiographs demonstrated extensive osteoarthritic changes involving the glenohumeral joint with a large inferior oriented osteophyte. Subsequent MRI of the shoulder was obtained which demonstrated isolated fatty atrophy of the teres minor and, to a lesser extent, deltoid muscles. The axillary nerve was visualized entering the quadrilateral space which, although, was severely narrowed secondary to the large osteophyte. The patient's clinical symptoms and MRI findings were consistent with quadrilateral space syndrome. The patient wanted to attempt conservative therapy first; and therefore, subsequently underwent physical therapy with improvement of shoulder strength and range of motion.