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The use of uncrewed systems within NOAA as a research and operational tool is well-documented. The full benefits of uncrewed systems to achieving the NOAA mission have remained largely unrealized, primarily because of the programmatic nature of these deployments. Establishment of enterprise services to coalesce uncrewed systems activities is essential for NOAA to optimize the use and benefits of these systems. A robust and encompassing data enterprise is principal among these needed enterprise services, and is essential to the success of NOAA's uncrewed systems strategy. The NOAA Uncrewed Systems Strategic Implementation Plan presents a comprehensive multi-year plan of action that builds upon the framework established in the NOAA Uncrewed Systems Strategy. This manuscript describes the Uncrewed Systems Data Enterprise as outlined in the Implementation Plan. In Section II we describe the data enterprise milestones. The notional operational framework and planned case studies are described in Sections III and IV, respectively. The advantages of the data enterprise are summarized in Section V.

Purpose: Antibodies that target immune checkpoint molecules have demonstrated significant and durable clinical benefit through re-activation and proliferation of pre-existing tumor-infiltrating CD8+ T cells in a broad range of tumor types including lung cancer. Detailed characterization of T cell dynamics at clonal levels using nextgeneration T cell receptor (TCR) sequencing in large cohorts of lung cancer patients is yet reported.Methods: We performed TCR sequencing of peripheral blood samples (and some tumors) obtained from 27 lung cancer patients undergoing single/combined immune checkpoint blockade therapies.Results: In one responder, we found expansion of a single T cell clone (approximately 20% of the all TCR reads) in a metastatic subcutaneous lesion that showed pathologic complete response on day 17 of treatment. The same TCR CDR3 sequence was detected in 6.1% of TCR reads in the peripheral blood prior to treatment initiation and the expansion remained persistent in peripheral blood at 21.0% at week 10 and 24.3% at week 48. In other patients who showed durable response, we also found persistent oligoclonal T cell expansion in their peripheral blood which was not observed in non-responders even while they remained on therapy.Conclusion: We found sustained expansion of oligoclonal T cell clones in lung cancer patients who had durable response to immune checkpoint blockade. This suggests possible use of longitudinal TCR sequencing to assist clinical decision-making, assess synergism with other agents, and most importantly facilitate rational development of alternative treatment strategies for non-responders.

This Environmental Assessment has been developed for the Replace Family Housing Phase 6 and Phase 7 design and construction project at Malmstrom AFB. The current family housing situation at Malmstrom AFB is very poor. Of the 1,471 housing units on base, only 498 are deemed adequate according to current Air Force Housing Guidance (USAF 1995a, 2003). Many of the existing homes have deteriorated and the following problems have been reported electrical wiring and fixtures that do not meet current building codes, plumbing fixtures that have corroded, outdated flooring, asbestos is present in flooring and countertops, and lead-based paint has been detected on several interior and exterior surfaces. The Air Force proposes to build new housing units within the existing housing area to remedy these needs.

This EA has been developed for the Replace Family Housing Phase V (Jupiter) design and construction project at Malmstrom AFB. The current family housing situation at Malmstrom AFB is very poor. Of the 1,406 housing units on base, only 377 are deemed adequate according to current AF Housing Guidance (USAF 1995a, 2003). Many of the existing homes have deterioirated and the following problems have been reported: electrical wiring and fixtures that do not meet current building codes, plumbing fixtures that have corroded, outdated flooring, asbestos is present in flooring and countertops, and lead-based paint has been detected on several interior and exterior surfaces. The AF proposes to build new housing units within the existing housing area to remedy these needs.

Ten discrete fractions of chondroitin 4-sulfate were prepared and refined by polyacrylamide gel electrophoresis. The molecular weights were determined based on the molar ratios of the components to the end group xylose. They have the same free electrophoretic mobility and the logarithms of their molecular weights were linearly related to the relative electrophoretic mobilities in the disc polyacrylamide gel electrophoresis. Using some of the fractions as standards, the polydispersity of the chondroitin sulfate was proven to be mainly due to a continuum of varying chain length.

Soft X-ray contact microscopy is a relatively new form of ultrastructural imaging, having better than 6 nm resolution and being uniquely well suited for the examination of fragile, unstained biological specimens. The biological specimen placed on a layer of photoresist and exposed to soft X-rays (1-10 nm lambda) of a specific wavelength or broad band. After X-ray exposure, the specimen is removed from the photoresist and the latter chemically developed. When the developed replica is examined by high resolution scanning electron microscopy, the fine structure of the original biological specimen is faithfully reproduced. Since the soft X-ray replica is initially formed due to the differential absorption of the incident X-rays by the biological specimen, the resultant contact replica also reveals information about the elemental composition of the sample. This paper presents our application of this new technique for the study of the proteoglycans, the complex polyanionic macromolecules comprising the gel phase in the matrix of mammalian cartilage.

During the past 10 years, attempts have been made to examine hydrated biological specimens by using wet chambers (at ambient temperature) [1-3] or cold stages (at -30 degrees C and below) during electron microscopic examination. Obtaining sufficient contrast of unstained hydrated biological samples has proven a considerable problem using both of these methods. Many fragile biological specimens, when examined hydrated, frozen or dried, are severely damaged by the electron beam and cannot be imaged by conventional scanning or transmission electron microscopy. In order to increase specimen contrast and eliminate electron beam induced trauma to the specimen, we have developed a wet-cell [4], which when used in concert with a pulsed plasma soft X-ray source, provides high contrast contact replicas of totally hydrated, unstained biological specimens. Although it has been postulated that hydrated unstained samples can be imaged by soft X-ray contact microscopy [5-7], to date there has been little success due to cell movement or degradation of the wet sample during the long exposure period necessary for an adequate imaging dose [8]. With the pulsed plasma source described in this study we have been able to use exposure times of approximately 40-60 ns while maintaining the sample in its hydrated state at atmospheric pressure. The resultant contact replicas exhibit good contrast and better than 30 nm spatial resolution when examined by conventional scanning electron microscopy.

Evidence is presented which establishes that chondroitin sulfate and keratan sulfate are part of the same macromolecular proteinpolysaccharide in bovine nasal septum. Degradation of the proteinpolysaccharide with papain releases single chains of chondroitin sulfate with a characteristic residual peptide still attached. A larger molecular weight fragment contains the keratan sulfate which retains a different distribution of amino acids and some chondroitin sulfate chains which can be separated by treatment with alkali with the loss of serine and theronine residues. While the ratio of protein to chondroitin sulfate varies widely in proteinpolysaccharide, the ratio of protein to keratan sulfate remains remarkably constant and different methods of degradation which release single chains of chondroitin sulfate from the macromolecule normally retain the keratan sulfate in the high molecular weight residue.

Proteinpolysaccharide can be extracted readily at 37° from bovine cartilage without destruction of the collagenous structure. The proteinpolysaccharide, regardless of the method of extraction, contains a variable ratio of chondroitin sulfate chains to protein as shown by preparative electrophoretic fractionation. More vigorous methods of extraction cause degradation of the proteinpolysaccharide and gel filtration distinguishes between single chains of chondroitin sulfate and multichain proteinpolysaccharide. Electrophoretic fractions which are richer in chondroitin sulfate chains show a corresponding increase in alkali-sensitive serine residues. These serines are not evenly distributed with respect to the core material. Other considerations make it unlikely that the sensitive serines are sequentially located in a core protein.

Summary1) Further examples of quantity and types of urinary excretion of sulfated mucopolysaccharides in Hurler's syndrome have been studied in 9 cases. All excreted both ChS-B as the major compone...

When high-density, cartilage proteoglycan (and the lower molecular weight mucopolysaccharides derived from it by papain or trypsin digestion) is centrifuged to equilibrium in complex gradients of CsCl and guanidinium chloride, the peak density is markedly and reproducibly decreased. Hyaluronic acid, a less dense macromolecule, is also decreased in density by the presence of guanidinium ion in the gradient and the extent of the decrease is directly related to the concentration of the guanidinium ion in the starting solution. Other cations, such as Li + or Ca 2+ , also produce marked, concentration-dependent, decreases in the density of hyaluronate. The effect is complicated by the fact that the equilibrium distribution of the added cation does not follow the CsCl gradient, but forms a reverse gradient. The density shift must involve an interaction of the added cation with the charged groups of the mucopolysaccharide polyanion since the added cations have no effect on uncharged polysaccharides such as dextran or Ficoll. Furthermore, it seems unlikely that the primary effect could be caused by increased hydration due to increased water activity since centrifugation in Cs 2 SO 4 (which reduces the density of nucleic acids by increasing their hydration) leads to substantial decreases in density of all polysaccharides including the uncharged dextran and Ficoll. Although the density of proteins, such as albumin and lysozyme, is not appreciably affected by guanidinium ion; proteoglycan and isolated keratan sulfate fractions of high protein content are markedly decreased in density by guanidinium ion. Such density shifts of the components of proteoglycan must be considered in any attempt to interpret experiments which study disaggregation phenomena in CsCl/guanidinium chloride gradients.

A systematic study has been made of the types and quantities of acid mucopolysaccharides isolated from a variety of tissues and fluids. Identification of the polysaccharides has been based on the precipitation of the polysaccharides as calcium salts, on analysis, on optical rotation and on digestibility by testicular and bacterial hyaluronidases. On this basis, six distinct mucopolysaccharides have been established: hyaluronic acid, chondroitin, the chondroitin sulfates designated as A, B and C, and keratosulfate. Additional mucopolysaccharides have been demonstrated in amyloid tissue and, in aorta, which are resistant to testicular hyaluronidase and contain D-glucosamine, a uronic acid and varying amounts of sulfate. In addition, incompletely sulfated chondroitin sulfates A or C have been obtained by fractional elution from an ion exchange resin. The tissues have been divided into seven groups on the basis of the mucopolysaccharides isolated from them, as follows: 1. (1) Vitreous humor, synovial fluid, filterable fowl tumors, liposarcoma: hyaluronic acid only. 2. (2) Cartilage, adult bone, chondrosarcoma, chordoma: chondroitin sulfate A and/or C. 3. (3) Umbilical cord, loose connective tissue of electric eel, fibroblasts grown in tissue culture: hyaluronic acid as the major with chondroitin sulfate C as a minor component. 4. (4) Pig skin, ligamentum nuchae (the latter contains chondroitin sulfate A in addition): chondroitin sulfate B and hyaluronic acid as major components. 5. (5) Adult connective tissue, tendon, heart valve: chondroitin sulfate B and C with hyaluronate as minor component. 6. (6) Cornea: keratosulfate, chondroitin and chondroitin sulfate A. 7. (7) Incompletely characterized fractions (amyloid, bovine aorta, calf bone). From calf bone, chondroitin sulfate A and C and a large incompletely sulfated fraction were isolated and, in addition, a minor fraction of keratosulfate. From bovine aorta, a major fraction with the properties of chondroitin sulfate A was isolated, accompanied by hyaluronate, chondroitin sulfate B and a variety of glucosamine-containing fractions of varying degrees of sulfation. These latter fractions are believed to be related to heparin. All normal tissues investigated appear to contain two or more mucopolysaccharides in contrast to tumors of mesodermal origin which produce a single type of mucopolysaccharide.

1. 1. ChS-B occurs in bullskin mainly in the middle and bottom layers. 2. 2. The fraction designated as ChS-B probably is a mixture of at least two isomeric mucopolysaccharides, the major component, the chondroitin sulfate B proper, containing iduronic acid, and the other containing glucuronic acid. Both are resistant to testicular hyaluronidase. 3. 3. The chemically desulfated product of chondroitin sulfate A was hydrolyzed by testicular and bacterial hyaluronidases analogous to their action on hyaluronate and chondroitin from cornea, while the desulfated products of chondroitin sulfate B remained resistant to the two hyaluronidases. 4. 4. Chondroitin sulfate B appears to be associated with the coarse bundle type of collagen fibers.

Two types of hyaluronidases have been described: one, found in testicular extracts and in snake venom, randomly cleaves the hexosaminidic bonds and yields a consecutive series from disaccharide through higher homologues of disaccharide units1; the other, found in micro-organisms, yields a Δ4–5 (glucuronido) unsaturated disaccharide2. In this communication, a third type of hyaluronidase is described, found thus far only in extracts of the leech (Hirudo medicinalis), which hydrolyses the endoglucuronidic linkages of hyaluronic acid.

Summary1. From the urine of 4 out of 5 patients with gargoylism, a mixture of mucopolysaccharides was isolated which could be identified as ChS-B and heparitin sulfate—with the former predominating. The urine of one patient, and the liver of another, yielded only heparitin sulfate. 2. The type of polysaccharide excreted by these patients had no obvious correlation with the severity of the disease or the postulated mode of inheritance suggested by the history. 3. It is suggested that gargoylism represents an overproduction of certain mucopolysaccharides due to a genetically determined error of differentiation of fibroblasts.