Optics research at the U.S. Army Research Laboratory.

The U.S. Army Research Laboratory (ARL) is the Army's premier laboratory for land forces. The Army relies on ARL for scientific discoveries, technological advances, and analyses that enable capabilities a future Army will need to persevere over adversaries. Although a relatively young organization that will celebrate 25 years of the discovery, innovation, and transition of science and technology in October 2017, ARL has already had significant impact in a wide range of scientific and technological disciplines. In this paper, we highlight some of its past and recent achievements in optics and photonics.

Newborn screening for X-linked adrenoleukodystrophy in New York State: diagnostic protocol, surveillance protocol and treatment guidelines.

PURPOSE: To describe a diagnostic protocol, surveillance and treatment guidelines, genetic counseling considerations and long-term follow-up data elements developed in preparation for X-linked adrenoleukodystrophy (X-ALD) newborn screening in New York State. METHODS: A group including the director from each regional NYS inherited metabolic disorder center, personnel from the NYS Newborn Screening Program, and others prepared a follow-up plan for X-ALD NBS. Over the months preceding the start of screening, a series of conference calls took place to develop and refine a complete newborn screening system from initial positive screen results to long-term follow-up. RESULTS: A diagnostic protocol was developed to determine for each newborn with a positive screen whether the final diagnosis is X-ALD, carrier of X-ALD, Zellweger spectrum disorder, acyl CoA oxidase deficiency or D-bifunctional protein deficiency. For asymptomatic males with X-ALD, surveillance protocols were developed for use at the time of diagnosis, during childhood and during adulthood. Considerations for timing of treatment of adrenal and cerebral disease were developed. CONCLUSION: Because New York was the first newborn screening laboratory to include X-ALD on its panel, and symptoms may not develop for years, long-term follow-up is needed to evaluate the presented guidelines.

Adenoviral transfer of human aquaporin-1 gene to rat liver improves bile flow in estrogen-induced cholestasis.

Estrogens can cause liver cholestatic disease. As downregulation of hepatocyte canalicular aquaporin-8 (AQP8) water channels has been involved in estrogen-induced bile secretory failure, we tested whether the archetypal water channel AQP1 improves 17α-ethinylestradiol (EE)-induced cholestasis. EE administration to rats reduced bile flow by 50%. A recombinant adenoviral (Ad) vector encoding human AQP1 (hAQP1), AdhAQP1, or a control vector was administered by retrograde bile ductal infusion. Hepatocyte canalicular hAQP1 expression was confirmed by liver immunostaining and immunoblotting in purified membrane fractions. Accordingly, canalicular osmotic water permeability was markedly increased. Bile flow, either basal or bile salt-stimulated was significantly augmented by over 50%. The choleretic efficiency of endogenous bile salts (that is, volume of bile per µmol of excreted bile salt) was significantly increased by 45% without changes in the biliary bile salt composition. Our data suggest that the adenoviral transfer of hAQP1 gene to the livers of EE-induced cholestatic rats improves bile flow by enhancing the AQP-mediated bile salt-induced canalicular water secretion. This novel finding might have potential therapeutic implications for cholestatic diseases.

Observation of suppression of light scattering induced by dipole-dipole interactions in a cold-atom ensemble.

We study the emergence of collective scattering in the presence of dipole-dipole interactions when we illuminate a cold cloud of rubidium atoms with a near-resonant and weak intensity laser. The size of the atomic sample is comparable to the wavelength of light. When we gradually increase the number of atoms from 1 to ~450, we observe a broadening of the line, a small redshift and, consistently with these, a strong suppression of the scattered light with respect to the noninteracting atom case. We compare our data to numerical simulations of the optical response, which include the internal level structure of the atoms.

Direct measurement of the Wigner time delay for the scattering of light by a single atom.

We have implemented the Gedanken experiment of an individual atom scattering a wave packet of near-resonant light, and measured the associated Wigner time delay as a function of the frequency of the light. In our apparatus, the atom behaves as a two-level system and we have found delays as large as 42 ns at resonance, limited by the lifetime of the excited state. This delay is an important parameter in the problem of collective near-resonant scattering by an ensemble of interacting particles, which is encountered in many areas of physics.

Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming.

During hypothermic preservation of cells (0-4°C), metabolism is diminished and energy-dependent transport processes are arrested. The effect of hypothermic preservation of hepatocytes in endocytic transport following rewarming has not been previously reported. We evaluated the uptake of EGF (Epidermal Growth Factor) ligand conjugated to fluorescent Quantum Dots (QDs) probes in rat hepatocytes after 24 and 72h cold storage in University of Wisconsin (UW) solution at 4°C. QDs uptake was visualized during rewarming to 37°C under air or, in a second approach, at the end of rewarming under 5% CO2. After 24h in UW solution, QDs were internalized under both rewarming conditions similar to non-preserved hepatocytes and cells maintained a normal cytoskeleton distribution. However, in hepatocytes preserved 72h none of the cells internalized QDs, which remained bound to the membranes. After rewarming, this group showed diminished actin staining and 60% reduction in ATP levels, while viability was maintained at ∼70%. Our results present evidence that, hypothermic preservation for 72h in UW solution at 4°C does not prevent EGFR (epidermal growth factor receptor) activation but irreversibly impairs endocytic uptake upon EGF stimulation; presumably due to actin cytoskeleton disassembling besides reduced ATP pool. Our approach can be applied on other membrane receptor systems and with other hypothermic preservation solutions to understand the effect of cooling in endocytic transport and to determine the optimal cold storage period.

Blended learning for accredited life support courses - A systematic review.

Aim: To evaluate the effectiveness on educational and resource outcomes of blended compared to non-blended learning approaches for participants undertaking accredited life support courses. Methods: This review was conducted in adherence with PRISMA standards. We searched EMBASE.com (including all journals listed in Medline), CINAHL and Cochrane from 1 January 2000 to 6 August 2021. Randomised and non-randomised studies were eligible for inclusion. Study screening, data extraction, risk of bias assessment (using RoB2 and ROBINS-I tools), and certainty of evidence evaluation (using GRADE) were all independently performed in duplicate. The systematic review was registered with PROSPERO (CRD42022274392). Results: From 2,420 studies, we included data from 23 studies covering fourteen basic life support (BLS) with 2,745 participants, eight advanced cardiac life support (ALS) with 33,579 participants, and one Advanced Trauma Life Support (ATLS) with 92 participants. Blended learning is at least as effective as non-blended learning for participant satisfaction, knowledge, skills, and attitudes. There is potential for cost reduction and eventual net profit in using blended learning despite high set up costs. The certainty of evidence was very low due to a high risk of bias and inconsistency. Heterogeneity across studies precluded any meta-analysis. Conclusion: Blended learning is at least as effective as non-blended learning for accredited BLS, ALS, and ATLS courses. Blended learning is associated with significant long term cost savings and thus provides a more efficient method of teaching. Further research is needed to investigate specific delivery methods and the effect of blended learning on other accredited life support courses.

Chemoprophylactic agent in schistosomiasis: 14,15-epoxygeranylgeraniol.

The occurrence of all-trans (-)-14,15-epoxygeranyl-geraniol in Pterodon pubescens Benth. is established, and its prophylactic activity against infection by Schitsosma mansoni demonstrated. Two other diterpenes present in the oil are inactive.

DNA array analysis for red blood cell antigens facilitates the transfusion support with antigen-matched blood in patients with sickle cell disease.

BACKGROUND: Blood samples from patients with sickle cell disease (SCD) present to transfusion service with numerous antibodies, making the searching for compatible red blood cells (RBC) a challenge. To overcome this problem we developed an effective strategy to meet needs of supplying RBC-compatible units to SCD patients using DNA arrays. METHODS: We selected DNA samples from 144 SCD patients with multiple (receiving > 5 units) transfusions previously phenotyped for ABO, Rh(D, C, c, E, e), K1, Fy(a) and Jk(a). We also selected DNA samples from 948 Brazilian blood donors whose ABO/RhD phenotype matched that of the patients. All samples were analysed by DNA array analysis (HEA Beadchip(TM), Bioarray Solutions) to determine polymorphisms associated with antigen expression for 11 blood group systems (Rh, Kell, Kidd, Duffy, MNS, Dombrock, Lutheran, Landsteiner-Wiener, Diego, Colton, Scianna); and one mutation associated with haemoglobinopathies. RESULTS: Based on genotype results we were able to predict phenotype-compatible donors needed in order to provide compatible units to this group of patients. Based on their ABO/Rh phenotype we were able to find in this pool of donors compatible units for 134 SCD patients. CONCLUSION: Blood group genotyping by DNA array contributes to the management of transfusions in SCD patients by facilitating the transfusion support with antigen-matched blood. It has the potential to improve the life of thousands of SCD-transfused patients by reducing mortality due to transfusion reactions and immunization.

Rapid generation of HNO induced by visible light.

We present a new method for controlled generation of HNO, based on the combination of a pH photoactuator induced by visible light with an HNO donor activated by pH increase. This method avoids the use of UV light, and in the future could be extended by using an IR photoactuator.

Taurolithocholate can inhibit the biliary discharge of lysosomes in the rat.

The natural bile salt taurolithocholate (TLC) impairs the biliary excretion of lipids and proteins, which are known to reach the canaliculus via vesicles. In this study we examined whether these observations could be extended to the exocytic discharge of lysosomal contents into bile. The single intravenous injection of a cholestatic dose of TLC, 3 micromol/100 g body wt., markedly inhibited the biliary excretion of the lysosomal enzymes acid phosphatase and beta-glucuronidase, despite the excretion of bile salts being normalized after a transient diminution. Under such a condition, TLC did not affect the normal transport to and the processing in lysosomes of the exogenously administered [14C]sucrose-labeled horseradish peroxidase. However, the biliary excretion of the radioactive lysosomal metabolites of the protein was significantly reduced. The results indicate that TLC can inhibit the biliary discharge of lysosomes in the rat without altering the functional integrity of these organelles. Possible explanations for these findings are discussed.

Taurolithocholate-induced inhibition of biliary lipid and protein excretion in the rat.

Taurolithocholate (TLC), a natural bile salt, induces selective impairment on canalicular membrane of the hepatocyte, which seems to be a major determinant of its cholestatic effect in experimental animals. In order to extend existing studies about the effects of TLC on bile secretion, we examined in TLC-treated rats the biliary excretion of compounds that are transported to canalicular membrane via vesicles, such as lipids and proteins. The single intravenous injection of TLC (3 mumol/100 g body wt.) inhibited transiently the biliary bile salt excretion, while the biliary excretion of lipids (i.e., cholesterol and phospholipids) and proteins remained inhibited even though the biliary excretion and composition of bile salts were normalized. Under such a condition, TLC also inhibited the transcellular vesicular pathway to the exogenous protein horseradish peroxidase entry into bile, without altering the paracellular biliary access of the protein. The hepatic uptake of horseradish peroxidase was unaffected by TLC-treatment. The results indicate that TLC can inhibit the biliary excretion of compounds that reach the canaliculus via a vesicular pathway, such as lipids and proteins, by a mechanism not related to a defective bile salt excretion. Possible explanations for these findings are discussed.

Hepatic handling of photoirradiated bilirubin. A study in isolated perfused Wistar rat liver.

Conjugation has been considered the rate-limiting step for bilirubin hepatic transport, and bypass of this metabolic step could explain why photobilirubins can be rapidly cleared by the liver. In this paper we assessed whether photoirradiation may enhance the bilirubin overall hepatic transport in the isolated perfused Wistar rat liver, a model possessing intact transport and conjugating systems. Bilirubin was administered as a bolus so as to reach a perfusate concentration of approximately 10 microM (bilirubin/albumin molar ratio 1:17). Perfusate light exposure (0.56.10(15) quanta s-1 cm-2) yielded 7-10% of configurational photoisomers, which were further identified as (4Z,15E/4E,15Z)-bilirubin IX alpha. Under such conditions, the perfusate removal rate was increased by 39% over that from dark conditions. Likewise, biliary excretion, estimated as total bilirubin recovery at 60 min, was also increased (+48%). This later improvement was mainly produced at the expense of unconjugated bilirubin, which most likely derived from its configurational photoisomers that, once excreted into bile, readily re-isomerized to the parent compound. In addition, this increment was partially due to a delayed improvement of monoglucuronide pigment excretion. The calculated hepatic pigment content was significantly higher under light conditions. A direct assessment of hepatic content of different bilirubin moieties at 20 min after bilirubin administration confirmed that such an increment was fully accounted for by unconjugated pigment. Our finding that hepatic pigment content rose (despite a higher biliary excretion) when the bilirubin was irradiated suggests a higher net uptake of photoisomers than native pigment. This observation, and the finding that bilirubin photoisomers were usually excreted without undergoing conjugation even if the metabolic system is active, contribute to explain the greater appearance of unconjugated bilirubin in Wistar rat bile under light exposure.

Taurocholate-induced inhibition of hepatic lysosomal degradation of horseradish peroxidase.

Endocytosed proteins in hepatocytes are transported to lysosomes for degradation. Metabolites accumulating in these organelles are released into bile by exocytosis, a process that seems to be regulated by the bile salt taurocholate (TC). In this study we examined if TC is also involved in the control of the lysosomal degradation of endocytosed proteins. We used [(14)C]sucrose-labeled horseradish peroxidase ([(14)C]S-HRP), a probe suitable to evaluate lysosomal proteolysis. TC-infused rats as well as isolated rat hepatocytes exposed to TC showed a significant inhibition in the lysosomal degradation of [(14)C]S-HRP (approximately 30%), with no change in either the uptake or the amount of protein reaching lysosomes. Under these conditions, the in vitro assay of lysosomal cathepsins B, L, H, and D revealed no change in their activities, suggesting that a reversible inhibition (lysosomal alkalinization?) was taking place in hepatocytes. Nevertheless, lysosomal pH measured using fluorescein isothiocyanate-dextran was shown not to be altered by TC. In addition, TC was unable to inhibit proteolysis in [(14)C]S-HRP loaded lysosomes or interfere in cathepsin assays. The results suggest that TC inhibits the lysosomal degradation of endocytosed proteins in hepatocytes and that the mechanism does not involve an effect of the bile salt per se or a rise in lysosomal pH.

Induction of phase II biotransformation reactions in rat jejunum during lactation. Possible involvement of prolactin.

The effect of lactation on UDP-glucuronosyltransferase (UGT) and Glutathione S-transferase (GST) activities was studied in jejunum from mother rats, 14 (LM14) and 21 (LM21) days after delivery. p-Nitrophenol glucuronidation rate was increased in LM14 and LM21 rats while conjugation of bilirubin and estrone was not affected and androsterone glucuronidation was decreased. Additional studies, including Western blotting and microsomal lipid analysis, revealed that the enhancement in p-nitrophenol UGT activity is most likely associated with an inductive process rather than with a modification in enzyme constraint. GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB) was also increased in LM14 and LM21 while activity towards 1,2-dichloro-4-nitrobenzene (DCNB) was not affected. Western blotting revealed a significant increase in the cytosolic content of mu (rGSTM2) and pi (rGSTP1) class subunits in LM14 and LM21 groups, while the alpha class subunit rGSTA2 remained unchanged. To evaluate the potential modulatory role of prolactin on the same enzyme systems, ovariectomized rats were treated with ovine prolactin (oPRL) at doses of 100, 200 and 300 microg/100 g body wt. per day for 4 days. Hormone administration affected UGT activities towards p-nitrophenol and androsterone and GST activity towards CDNB in a way and magnitude consistent with those produced in lactating rats, while conjugation of estrone, bilirubin and DCNB were unchanged. Western blotting data were also consistent with those of lactating rats. These results indicate that UGT and GST activities are increased in rat jejunum during lactation, due to induction of some specific isoforms, and that prolactin is the likely mediator of these effects.

Predicting membrane flux decline from complex mixtures using flow-field flow fractionation measurements and semi-empirical theory.

Flow-Field Flow Fractionation (FI-FFF) is an idealization of the cross flow membrane filtration process in that, (1) the filtration flux and crossflow velocity are constant from beginning to end of the device, (2) the process is a relatively well-defined laminar-flow hydrodynamic condition, and (3) the solutes are introduced as a pulse-input that spreads due to interactions with each other and the membrane in the dilute-solution limit. We have investigated the potential for relating FI-FFF measurements to membrane fouling. An advection-dispersion transport model was used to provide 'ideal' (defined as spherical, non-interacting solutes) solute residence time distributions (RTDs) for comparison with 'real' RTDs obtained experimentally at different cross-field velocities and solution ionic strength. An RTD moment analysis based on a particle diameter probability density function was used to extract "effective" characteristic properties, rather than uniquely defined characteristics, of the standard solute mixture. A semi-empirical unsteady-state, flux decline model was developed that uses solute property parameters. Three modes of flux decline are included: (1) concentration polarization, (2) cake buildup, and (3) adsorption on/in pores, We have used this model to test the hypothesis-that an analysis of a residence time distribution using FI-FFF can describe 'effective' solute properties or indices that can be related to membrane flux decline in crossflow membrane filtration. Constant flux filtration studies included the changes of transport hydrodynamics (solvent flux to solute back diffusion (J/k) ratios), solution ionic strength, and feed water composition for filtration using a regenerated cellulose ultrafiltration membrane. Tests of the modeling hypothesis were compared with experimental results from the filtration measurements using several correction parameters based on the mean and variance of the solute RTDs. The corrections used to modify the boundary layer mass transfer coefficient and the specific resistance of cake or adsorption layers demonstrated that RTD analysis is potentially useful technique to describe colloid properties but requires improvements.

Enhanced transport and liquid membranes in bioseparations.

Membranes that use a reversible chemical reaction or sequestration to achieve high selectivities and productivities show great potential for use in bioseparations. The concept of liquid membranes, with and without a complexing agent (carrier), and the types of system configurations and carriers that may be used with these membranes, are discussed.

Mapping of hereditary neuralgic amyotrophy (familial brachial plexus neuropathy) to distal chromosome 17q.

Hereditary neuralgic amyotrophy with predilection for the brachial plexus (HNA) is an autosomal dominant disorder associated with recurrent, episodic, painful brachial neuropathies. Mildly dysmorphic facial features, including hypotelorism, long nasal bridge, and upslanting palpebral fissures, are present in affected persons in some pedigrees with HNA. To determine the chromosomal location of the HNA gene, we carried out genetic linkage studies with polymerase chain reaction-based DNA markers in two large pedigrees. Linkage to markers from the distal long arm of chromosome 17 was established.

Effect of spironolactone on bilirubin metabolism in rat liver and small intestinal mucosa.

In vitro and in vivo experimental models were designed for the study of the effect of spironolactone (SP) on bilirubin metabolism in rat liver and small intestinal mucosa. In vitro studies included uptake of bilirubin by liver slices and intestinal sheets, determination of glucuronyltransferase activity in mucosal homogenates, and the handling of bilirubin by the isolated perfused liver after bilirubin overload. In vitro studies were carried out to measure the plasma disappearance rate of bilirubin and to determine the extent of bilirubin conjugation and biliary excretion of the pigment infused intravenously. The results obtained suggested that the mechanisms involved in the uptake of bilirubin by tissues were not influenced by SP pretreatment. Glucuronyltransferase activity in the small intestinal mucosa was significantly induced by SP, as previously observed in rat liver. Isolated perfused livers from SP-treated rats, as well as treated living rats, exhibited a greater than normal capacity for bilirubin excretion into bile at the expense of bilirubin diglucuronide. Conjugated bilirubin in the small intestinal mucosa of rats infused with unconjugated pigment was also increased after SP pretreatment. The results favoured the conclusion that SP is an inducer of bilirubin conjugation in the livers as well as in extrahepatic tissues, such as the small intestinal mucosa.

Effect of silymarin on biliary bile salt secretion in the rat.

The effect of the hepatoprotector silymarin on bile secretion, with particular regard to bile salt secretion, was studied in Wistar rats. Silymarin (25, 50, 100, and 150 mg/kg/day, i.p., for 5 days) induced a dose-dependent increase in bile flow and bile salt secretion, the maximal effect being reached at a dose of 100 mg/kg/day (+17 and +49%, for bile flow and bile salt output, respectively; P < 0.05). Assessment of bile salt composition in bile revealed that stimulation of the bile salt secretion was accounted for mainly by an increase in the biliary secretion of beta-muricholate and, to a lesser extent, of alpha-muricholate, chenodeoxycholate, ursodeoxycholate, and deoxycholate. The maximum secretory rate (T(m)) of bile salts, as assessed by infusing the non-hepatotoxic bile salt tauroursodeoxycholate i.v. at stepwise-increasing rates, was not influenced by silymarin. The flavonolignan also increased the endogenous bile salt pool size (+53%, P < 0.05) and biliary bile acid excretion after bile acid pool depletion (+54%, P < 0.05), a measure of de novo bile salt synthesis. These results suggest that silymarin increases the biliary excretion and the endogenous pool of bile salts by stimulating the synthesis, among others, of hepatoprotective bile salts, such as beta-muricholate and ursodeoxycholate.