503 resultados para Mandated reporters


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Background: Though 293T cells are widely used for expression of proteins from transfected plasmid vectors, the molecular basis for the high-level expression is yet to be understood. We recently identified the prostate carcinoma cell line PC3 to be as efficient as 293T in protein expression. This study was undertaken to decipher the molecular basis of high-level expression in these two cell lines. Methodology/Principal Findings: In a survey of different cell lines for efficient expression of platelet-derived growth factor-B (PDGF-B), beta-galactosidase (beta-gal) and green fluorescent protein (GFP) from plasmid vectors, PC3 was found to express at 5-50-fold higher levels compared to the bone metastatic prostate carcinoma cell line PC3BM and many other cell lines. Further, the efficiency of transfection and level of expression of the reporters in PC3 were comparable to that in 293T. Comparative analyses revealed that the high level expression of the reporters in the two cell lines was due to increased translational efficiency. While phosphatidic acid (PA)-mediated activation of mTOR, as revealed by drastic reduction in reporter expression by n-butanol, primarily contributed to the high level expression in PC3, multiple pathways involving PA, PI3K/Akt and ERK1/2 appear to contribute to the abundant reporter expression in 293T. Thus the extent of translational upregulation attained through the concerted activation of mTOR by multiple pathways in 293T could be achieved through its activation primarily by the PA pathway in PC3. Conclusions/Significance: Our studies reveal that the high-level expression of proteins from plasmid vectors is effected by translational up-regulation through mTOR activation via different signaling pathways in the two cell lines and that PC3 is as efficient as 293T for recombinant protein expression. Further, PC3 offers an advantage in that the level of expression of the protein can be regulated by simple addition of n-butanol to the culture medium.

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The high level of public accountability attached to Public Sector Enterprises as a result of public ownership makes them socially responsible. The Committee of Public Undertakings in 1992 examined the issue relating to social obligations of Central Public Sector Enterprises and observed that ``being part of the `State', every Public Sector enterprise has a moral responsibility to play an active role in discharging the social obligations endowed on a welfare state, subject to the financial health of the enterprise''. It issued the Corporate Social Responsibility Guidelines in 2010 where all Central Public Enterprises, through a Board Resolution, are mandated to create a CSR budget as a specified percentage of net profit of the previous year. This paper examines the CSR activities of the biggest engineering public sector organization in India, Bharath Heavy Electricals Limited. The objectives are twofold, one, to develop a case study of the organization about the funds allocated and utilized for various CSR activities, and two, to examine its status with regard to other organizations, the 2010 guidelines, and the local socio-economic development. Secondary data analysis results show three interesting trends. One, it reveals increasing organizational social orientation with the formal guidelines in place. Two, Firms can no longer continue to exploit environmental resources and escape from their responsibilities by acting separate entities regardless of the interest of the society and Three the thrust of CSR in public sector is on inclusive growth, sustainable development and capacity building with due attention to the socio-economic needs of the neglected and marginalized sections of the society.

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Unmet clinical needs remain the primary driving force for innovations in medical devices. While appropriate mechanisms to protect these innovative outcomes are essential, the performance of clinical trials to ensure safety is also mandated before the invention is ready for public use. Literature explaining the relationship between patenting activities and clinical trials of medical devices is scarce. Linking patent ownership to clinical trials may imply product leadership and value chain control. In this paper, we use patent data from Indian Patent Office (IPO), PCT, and data from Clinical Trials Registry of India (CTRI) to identify whether patent assignees have any role in leading as primary sponsors of clinical trials. A total of 42 primary sponsors are identified from the CTRI database in India. Number of patents awarded to these primary sponsors in the particular medical device, total number of patents awarded to the primary sponsor in all technologies, total number of patents in the specific medical device technology provides an indication of leadership and control in the value chain.

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3,4-Dichlorophenol (1) crystallizes in the tetragonal space group I4(1)/a with a short axis of 3.7926 (9) angstrom. The structure is unique in that both type I and type II Cl.....Cl interactions are present, these contact types being distinguished by the angle ranges of the respective C-Cl....Cl angles. The present study shows that these two types of contacts are utterly different. The crystal structures of 4-bromo-3-chlorophenol (2) and 3-bromo-4-chlorophenol (3) have been determined. The crystal structure of (2) is isomorphous to that of (1) with the Br atom in the 4-position participating in a type II interaction. However, the monoclinic P2(1)/c packing of compound (3) is different; while the structure still has O-H....O hydrogen bonds, the tetramer O-H.....O synthon seen in (1) and (2) is not seen. Rather than a type I Br....Br interaction which would have been mandated if (3) were isomorphous to (1) and (2), Br forms a Br....O contact wherein its electrophilic character is clearly evident. Crystal structures of the related compounds 4-chloro-3-iodophenol (4) and 3,5-dibromophenol (5) were also determined. A computational survey of the structural landscape was undertaken for (1), (2) and (3), using a crystal structure prediction protocol in space groups P2(1)/c and I4(1)/a with the COMPASS26 force field. While both tetragonal and monoclinic structures are energetically reasonable for all compounds, the fact that (3) takes the latter structure indicates that Br prefers type II over type I contacts. In order to differentiate further between type I and type II halogen contacts, which being chemically distinct are expected to have different distance fall-off properties, a variable-temperature crystallography study was performed on compounds (1), (2) and (4). Length variations with temperature are greater for type II contacts compared with type I. The type II Br....Br interaction in (2) is stronger than the corresponding type II Cl....Cl interaction in (1), leading to elastic bending of the former upon application of mechanical stress, which contrasts with the plastic deformation of (1). The observation of elastic deformation in (2) is noteworthy; in that it finds an explanation based on the strengths of the respective halogen bonds, it could also be taken as a good starting model for future property design. Cl/Br isostructurality is studied with the Cambridge Structural Database and it is indicated that this isostructurality is based on shape and size similarity of Cl and Br, rather than arising from any chemical resemblance.

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This report responds to the 1986 Beaches Bill which, in recognition of the potential deleterious impact on Florida's beaches of inlets modified for navigation, mandated a study of those inlets with identification of recommended action to reduce the impacts. This report addresses west Coast inlets; East Coast inlets are the subject of a companion report. There are 37 inlets along that portion of Florida's West Coast commencing from Pensacola Bay Entrance to Caxambas Pass at the south end of Marco Island. Compared to those on the East Coast, most West Coast inlets have not had the deleterious effects on the adjacent beaches, yet all modified inlets without proper management have the potential of impacting unfavorably on the adjacent shorelines. Moreover, at present there is interest in opening three West Coast entrances which either have been open in the past (Midnight Pass) or which have opened occasionally (Navarre Pass and Entrance to Phillips Lake). A review of inlets in their natural condition demonstrates the presence of a shallow broad outer bar across which the longshore transport Occurs. These shallow and shifting bar features were unsuitable for navigation which in many cases has led to the deepening of the channels and fixing with one or two jetty structures. Inlets in this modified state along with inappropriate maintenance practices have the potential of placing great ero$ional stress along the adjacent beaches. Moreover. channel dredging can reduce wave sheltering of the shoreline by ebb tidal shoals and alter the equilibrium of the affected shoreline segments. The ultimate in poor sand management practice is the placement of good quality beach sand in water depths too great for the sand to reenter the longshore system under natural forces; depths of 12 ft. or less are considered appropriate for Florida in order to maintain the sand in the system. With the interference of the nearshore sediment transport processes by inlets modified for navigation, if the adjacent beaches are to be stabilized there must be an active monitoring program with commitment to placement of dredged material of beach quality on shoreline segments of documented need. Several East Coast inlets have such transfer facilities; however. the quantities of sand transferred should be increased. Although an evolution and improvement in the technical capability to manage sand resources in the vicinity of inlets is expected, an adequate capability exists today and a concerted program should be made to commence a scheduled implementation of this capability at those entrances causing greatest erosional stress on the adjacent shorelines. A brief summary review for each of the 37 West Coast inlets is presented including: a scaled aerial photograph, brief historical information, several items related to sediment losses at each inlet and special characteristics relevant to State responsibilities. For each inlet, where appropriate, the above infor~tion is utilized to develop a recommenced action. (PDF has 101 pages.)

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Without knowledge of basic seafloor characteristics, the ability to address any number of critical marine and/or coastal management issues is diminished. For example, management and conservation of essential fish habitat (EFH), a requirement mandated by federally guided fishery management plans (FMPs), requires among other things a description of habitats for federally managed species. Although the list of attributes important to habitat are numerous, the ability to efficiently and effectively describe many, and especially at the scales required, does not exist with the tools currently available. However, several characteristics of seafloor morphology are readily obtainable at multiple scales and can serve as useful descriptors of habitat. Recent advancements in acoustic technology, such as multibeam echosounding (MBES), can provide remote indication of surficial sediment properties such as texture, hardness, or roughness, and further permit highly detailed renderings of seafloor morphology. With acoustic-based surveys providing a relatively efficient method for data acquisition, there exists a need for efficient and reproducible automated segmentation routines to process the data. Using MBES data collected by the Olympic Coast National Marine Sanctuary (OCNMS), and through a contracted seafloor survey, we expanded on the techniques of Cutter et al. (2003) to describe an objective repeatable process that uses parameterized local Fourier histogram (LFH) texture features to automate segmentation of surficial sediments from acoustic imagery using a maximum likelihood decision rule. Sonar signatures and classification performance were evaluated using video imagery obtained from a towed camera sled. Segmented raster images were converted to polygon features and attributed using a hierarchical deep-water marine benthic classification scheme (Greene et al. 1999) for use in a geographical information system (GIS). (PDF contains 41 pages.)

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The Olympic Coast National Marine Sanctuary (OCNMS or Sanctuary) planned and organized the 1998 Research Workshop as part of its mission to protect and improve understanding of its marine resources through research and education programs. The Sanctuary is also mandated to coordinate and facilitate information exchanges and sponsors periodic research workshops to that end. The goals of the 1998 Research Workshop were as follows: A. Highlight and prioritize research needs for the Sanctuary relative to the development of a framework for a five-year research plan; B. Build on results from the Olympic Coast Marine Research Workshop of 1996; C. Present recent/ongoing research; D. Share multi-disciplinary information; E. Select priority sites for multi-disciplinary studies; and F. Promote student participation and research. (PDF contains 93 pages.)

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This report reviews marine zoning in the Monterey Bay National Marine Sanctuary (MBNMS). The 72 zoned areas in the MBNMS are of 13 different zone types. Each marine zone type has associated regulations that restrict or promote specific activities. For example, recreational activities such as boating, fishing, tidepooling, snorkeling, and SCUBA diving are limited in some zones. Scientific research is allowed at all sites, with appropriate permits, and is specifically promoted in a few sites. In addition, motorized personal watercraft use, dredge material disposal, large vessel traffic, jade collection, and aircraft overflight are allowed only in specific zones. The effectiveness of the marine zoning in the MBNMS is difficult to determine for two reasons. Firstly, many of the zones lack a clearly stated purpose or have confusing regulations. Secondly, the majority of the zones have not been evaluated formally by the managing agencies. Of the zones that have been evaluated, such as Dredge Material Disposal zones, Big Creek MRPA Ecological Reserve, and Pt. Lobos State/Ecological Reserve, the majority appear to be achieving their mandated purpose to some extent. Many of the zones in the MBNMS fall under the title "marine reserve." Marine reserves have recently received significant attention internationally, nationally, and in California due to their potential for: improving the status of exploited species; protecting marine habitats and ecosystems from degradation; facilitating scientific research and fisheries management; and increasing ecotourism. However, reserves must be well designed and managed to reach this potential. A well designed and managed reserve will have clearly defined goals, scientifically-based design, proper enforcement of regulations, rigorous evaluation of the reserve's effectiveness, and adaptive management. Based on these criteria, the majority of the marine reserves in California are not well designed or managed. However, the State of California has recognized this problem and is in the process of re-evaluating the California system of marine managed areas. (PDF contains 137 pages.)

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In July 1974, we began a two-year baseline study of the Moss Landing Elkhorn Slough marine environment for Pacific Gas and Electric Company as mandated by the Coastal Commission. The original proposal included strong recommendations for more complete oceanographic studies and a third year of data collection. These further studies were not funded. This report is divided into three sections: oceanography, benthic invertebrate ecology and fish and zooplankton ecology. (PDF contains 480 pages)

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In July, 1974 we began a baseline study of the Moss Landing-Elkhorn Slough marine environment for PG&E as mandated by the Coastal Commission. This report constitutes results of the first year's program. It is divided into three sections, oceanography, benthic invertebrate ecology, and fish and zooplankton ecology. (PDF contains 226 pages)

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Shellfish bed closures along the North Carolina coast have increased over the years seemingly concurrent with increases in population (Mallin 2000). More and faster flowing storm water has come to mean more bacteria, and fecal indicator bacterial (FIB) standards for shellfish harvesting are often exceeded when no source of contamination is readily apparent (Kator and Rhodes, 1994). Could management reduce bacterial loads if the source of the bacteria where known? Several potentially useful methods for differentiating human versus animal pollution sources have emerged including Ribotyping and Multiple Antibiotic Resistance (MAR) (US EPA, 2005). Total Maximum Daily Load (TMDL) studies on bacterial sources have been conducted for streams in NC mountain and Piedmont areas (U.S. EPA, 1991 and 2005) and are likely to be mandated for coastal waters. TMDL analysis estimates allowable pollutant loads and allocates them to known sources so management actions may be taken to restore water to its intended uses (U.S. EPA, 1991 and 2005). This project sought first to quantify and compare fecal contamination levels for three different types of land use on the coast, and second, to apply MAR and ribotyping techniques and assess their effectiveness for indentifying bacterial sources. Third, results from these studies would be applied to one watershed to develop a case study coastal TMDL. All three watershed study areas are within Carteret County, North Carolina. Jumping Run Creek and Pettiford Creek are within the White Oak River Basin management unit whereas the South River falls within the Neuse River Basin. Jumping Run Creek watershed encompasses approximately 320 ha. Its watershed was a dense, coastal pocosin on sandy, relic dune ridges, but current land uses are primarily medium density residential. Pettiford Creek is in the Croatan National Forest, is 1133 ha. and is basically undeveloped. The third study area is on Open Grounds Farm in the South River watershed. Half of the 630 ha. watershed is under cultivation with most under active water control (flashboard risers). The remaining portion is forested silviculture.(PDF contains 4 pages)

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The relentlessly increasing demand for network bandwidth, driven primarily by Internet-based services such as mobile computing, cloud storage and video-on-demand, calls for more efficient utilization of the available communication spectrum, as that afforded by the resurging DSP-powered coherent optical communications. Encoding information in the phase of the optical carrier, using multilevel phase modulationformats, and employing coherent detection at the receiver allows for enhanced spectral efficiency and thus enables increased network capacity. The distributed feedback semiconductor laser (DFB) has served as the near exclusive light source powering the fiber optic, long-haul network for over 30 years. The transition to coherent communication systems is pushing the DFB laser to the limits of its abilities. This is due to its limited temporal coherence that directly translates into the number of different phases that can be imparted to a single optical pulse and thus to the data capacity. Temporal coherence, most commonly quantified in the spectral linewidth Δν, is limited by phase noise, result of quantum-mandated spontaneous emission of photons due to random recombination of carriers in the active region of the laser.

In this work we develop a generically new type of semiconductor laser with the requisite coherence properties. We demonstrate electrically driven lasers characterized by a quantum noise-limited spectral linewidth as low as 18 kHz. This narrow linewidth is result of a fundamentally new laser design philosophy that separates the functions of photon generation and storage and is enabled by a hybrid Si/III-V integration platform. Photons generated in the active region of the III-V material are readily stored away in the low loss Si that hosts the bulk of the laser field, thereby enabling high-Q photon storage. The storage of a large number of coherent quanta acts as an optical flywheel, which by its inertia reduces the effect of the spontaneous emission-mandated phase perturbations on the laser field, while the enhanced photon lifetime effectively reduces the emission rate of incoherent quanta into the lasing mode. Narrow linewidths are obtained over a wavelength bandwidth spanning the entire optical communication C-band (1530-1575nm) at only a fraction of the input power required by conventional DFB lasers. The results presented in this thesis hold great promise for the large scale integration of lithographically tuned, high-coherence laser arrays for use in coherent communications, that will enable Tb/s-scale data capacities.

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The aromatic core of double helical DNA possesses the unique and remarkable ability to form a conduit for electrons to travel over exceptionally long molecular distances. This core of π-stacked nucleobases creates an efficient pathway for charge transfer to proceed that is exquisitely sensitive to even subtle perturbations. Ground state electrochemistry of DNA-modified electrodes has been one of the major techniques used both to investigate and to harness the property of DNA-mediated charge transfer. DNA-modified electrodes have been an essential tool for both gaining insights into the fundamental properties of DNA and, due to the exquisite specificity of DNA-mediated charge transfer for the integrity of the π-stack, for use in next generation diagnostic sensing. Here, multiplexed DNA-modified electrodes are used to (i) gain new insights on the electrochemical coupling of metalloproteins to the DNA π-stack with relevance to the fundaments of in vivo DNA-mediated charge transfer and (ii) enhance the overall sensitivity of DNA-mediated reduction for use in the detection of low abundance diagnostic targets.

First, Methylene Blue (MB′) was covalently attached to DNA through a flexible C12 alkyl linker to yield a new redox reporter for DNA electrochemistry measurements with enhanced sensitivity. Tethered, intercalated MB′ was reduced through DNA-mediated charge transport. The redox signal intensity for MB′-dT-C12-DNA was found to be at least 3 fold larger than that of previously used Nile Blue (NB)-dT-DNA, which is coupled to the base stack via direct conjugation. The signal attenuation, due to an intervening mismatch, and therefore the degree of DNA-mediated reduction, does, however, depend on the DNA film morphology and the backfilling agent used to passivate the surface. These results highlight two possible mechanisms for the reduction of MB′ on the DNA-modified electrode that are distinguishable by their kinetics: reduction mediated by the DNA base pair stack and direct surface reduction of MB′ at the electrode. The extent of direct reduction at the surface can be minimized by overall DNA assembly conditions.

Next, a series of intercalation-based DNA-mediated electrochemical reporters were developed, using a flexible alkane linkage to validate and explore their DNA-mediated reduction. The general mechanism for the reduction of distally bound redox active species, covalently tethered to DNA through flexible alkyl linkages, was established to be an intraduplex DNA-mediated pathway. MB, NB, and anthraquinone were covalently tethered to DNA with three different covalent linkages. The extent of electronic coupling of the reporter was shown to correlate with the DNA binding affinity of the redox active species, supporting an intercalative mechanism. These electrochemical signals were shown to be exceptionally sensitive to a single intervening π-stack perturbation, an AC mismatch, in a densely packed DNA monolayer, which further supports that the reduction is DNA-mediated. Finally, this DNA-mediated reduction of MB occurs primarily via intra- rather than inter duplex intercalation, as probed through varying the proximity and integrity of the neighboring duplex DNA. Further gains to electrochemical sensitivity of our DNA-modified devices were then achieved through the application of electrocatalytic signal amplification using these solvent accessible intercalative reporters, MB-dT-C8, and hemoglobin as a novel electron sink. Electrocatalysis offers an excellent means of electrochemical signal amplification, yet in DNA based sensors, its application has been limited due to strict assembly conditions. We describe the use of hemoglobin as a robust and effective electron sink for electrocatalysis in DNA sensing on low density DNA films. Protein shielding of the heme redox center minimizes direct reduction at the electrode surface and permits assays on low density DNA films. Electrocatalysis of MB that is covalently tethered to the DNA by a flexible alkyl linkage allows for efficient interactions with both the base stack and hemoglobin. Consistent suppression of the redox signal upon incorporation of single CA mismatch in the DNA oligomer demonstrates that both the unamplified and the electrocatalytically amplified redox signals are generated through DNA-mediated charge transport. Electrocatalysis with hemoglobin is robust: it is stable to pH and temperature variations. The utility and applicability of electrocatalysis with hemoglobin is demonstrated through restriction enzyme detection, and an enhancement in sensitivity permits femtomole DNA sampling.

Finally, we expanded the application of our multiplexed DNA-modified electrodes to the electrochemical characterization of DNA-bound proteins containing [4Fe-4S] clusters. DNA-modified electrodes have become an essential tool for the characterization of the redox chemistry of DNA repair proteins that contain redox cofactors. Multiplexed analysis of EndonucleaseIII (EndoIII), a DNA repair protein containing a [4Fe-4S] cluster known to be accessible via DNA-mediated charge transport, elucidated subtle differences in the electrochemical behavior as a function of DNA morphology. DNA-bound EndoIII is seen to have two different electron transfer pathways for reduction, either through the DNA base stack or through direct surface reduction. Closely packed DNA films, where the protein has limited surface accessibility, produce electrochemical signals reflecting electron transfer that is DNA-mediated. The electrochemical comparison of EndoIII mutants, including a new family of mutations altering the electrostatics surrounding the [4Fe-4S] cluster, was able to be quantitatively performed. While little change in the midpoint potential was found for this family of mutants, significant variations in the efficiency of DNA-mediated electron transfer were apparent. Based on the stability of these proteins, examined by circular dichroism, we propose that the electron transfer pathway can be perturbed not only by the removal of aromatic residues, but also through changes in solvation near the cluster.

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O campo de estudos da Comunicação é conhecido pela sua interdisciplinaridade, o que permite analisar objetos relacionados a outras áreas do conhecimento por meio da perspectiva em que os diferentes tipos de mídia lhes apresentam. Utilizando palavras, imagens e sons os meios de comunicação colaboram para a produção dos significados de uso e apropriação de diferentes produtos e serviços disponíveis para o consumo. Por meio de um intercâmbio com o campo de estudos de consumo e tratando de um produto oriundo do campo da Educação, os cursos de MBA, este trabalho analisa as mensagens produzidas pelo Boa Chance, suplemento publicado pelo jornal O Globo. Para compor o referencial foram considerados teóricos do campo da Comunicação, Consumo, Educação e Representação Social. Além de oferecerem subsídios para refletir sobre o consumo desse tipo de produto, as idéias e conceitos apresentados também colaboram para compor as categorias elaboradas para a análise das matérias publicadas. Para aprofundar um pouco mais sobre os cursos de MBA foi traçado um histórico desse tipo de ensino, desde o seu surgimento nos Estados Unidos, passando pela sua ascensão a partir da década de 1940 até a sua chegada ao Brasil. A forma como os cursos adentraram a pauta dos cadernos de emprego, em especial, do Boa Chance, também é abordada. Entrevistas com repórteres e editores que trabalharam ou que ainda estão no caderno permitiram identificar o contexto em que as pautas relacionadas a esse tema se tornaram mais frequentes. A partir disso, foi possível compreender que o assunto começou a ganhar mais espaço na mesma medida que cresce a oferta de vagas pelas instituições, no começo dos anos 2000. Para entender de que maneira os meios de comunicação abordam para esse tipo de consumo e também quais são as representações que ele produz sobre os MBAs, foram analisadas 26 edições do caderno recolhidas entre 2009 e 2010.

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Horseshoe crabs (Limulus polyphemus) are valued by many stakeholders, including the commercial fishing industry, biomedical companies, and environmental interest groups. We designed a study to test the accuracy of the conversion factors that were used by NOAA Fisheries and state agencies to estimate horseshoe crab landings before mandatory reporting that began in 1998. Our results indicate that the NOAA Fisheries conversion factor consistently overestimates the weight of male horseshoe crabs, particularly those from New England populations. Because of the inaccuracy of this and other conversion factors, states are now mandated to report the number (not biomass) and sex of landed horseshoe crabs. However, accurate estimates of biomass are still necessary for use in prediction models that are being developed to better manage the horseshoe crab fishery. We recommend that managers use the conversion factors presented in this study to convert current landing data from numbers to biomass of harvested horseshoe crabs for future assessments.