Systemic and mucosal antibody responses specific to SARS-CoV-2 during mild versus severe COVID-19

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A functionally active complement system is present in uterine secretion of the mouse prior to implantation.

Sephadex beads were placed carefully in the uterus on days 2 and 3 and left for 6 to 8 h to absorb uterine secretion. The beads were then removed with volatile silicon oil and mounted on small pieces of nitrocellulose paper. Immuno-staining of these bead blots showed they contained the complement components C1q, C3, C4, and C5. We demonstrated that complement component C3 in the uterine secretion could be activated and deposited on model immune complexes, and also that antibody-coated erythrocytes were lysed in utero, that is, a membrane attack complex was produced. Thus, the mouse uterine secretion at the preimplantation stage contains a functionally active complement system.     

Threshold effect of urinary glycosaminoglycans and the walk test as indicators of disease progression in a survey of subjects with Mucopolysaccharidosis VI (Maroteaux-Lamy syndrome)

A cross-sectional survey in individuals affected with the lysosomal storage disease Mucopolysaccharidosis VI (MPS VI) was conducted to establish demographics, urinary glycosaminoglycan (GAG) levels, and clinical progression of the disease. The survey evaluated 121 bona fide MPS VI-affected individuals over the age of 4 years from 15 countries across the Americas, Europe, and Australasia representing greater than 10% of the estimated world prevalence of the disease. A medical history, complete physical exam, urinary GAG determination, and assessment of several clinical measures related to physical endurance, pulmonary function, joint range of motion, strength, and quality of life were completed for each participant. Although a wide variation in clinical presentation was observed, several general findings were obtained reflecting progression of the disease. Impaired physical endurance, as measured by the distance achieved in a 6-min walk, could be demonstrated across all age groups of MPS VI-affected individuals. High urinary GAG values (>200 mug/mg creatinine) were associated with an accelerated clinical course comprised of age-adjusted short stature and low body weight, impaired endurance, compromised pulmonary function, and reduced joint range of motion. An unexpected result was the predominance of urinary GAG values <100 mug/mg creatinine for those participants over the age of 20 years. Pending the collection of longitudinal data, these results suggest that urinary GAG levels predict clinical morbidity, and longer-term survival is associated with urinary GAG levels below a threshold of 100 mug/mg creatinine.     

Activity of Pilomotor Muscles of Single Tactile Hairs in the Cat

Contractions of the pilomotor muscles of individual carpal tactile sinus hairs in the cat were studied by simultaneous recording of the movement of a single hair by means of a capacitance meter, and of the electrical activity during the muscle contraction, as recorded by an external metal microelectrode. Single shock stimulation of the nerve gave rise to a twitch contraction, whereas repetitive stimulation caused summation of contractions at frequencies higher than 0.2/sec and fusion at frequencies of 1/sec or more. The electrical response consisted of a slow potential and a superimposed spike. These components were both shown to be dependent on the stimulus strength and to be facilitated on repetitive stimulation. At stimulus frequencies higher than 3/sec the spike gradually declined in amplitude and was substituted by a rhythmic oscillation at the same rate as the stimulus. Even when no spike was recorded the contraction was sustained. Comparisons are made between the electrical activity elicited in the pilomotor muscles and the junction potentials and spikes recorded in other types of smooth muscle.     

Comparative dosimetry of diode and diamond detectors in electron beams for intraoperative radiation therapy

The aim of the present study is to examine the validity of using silicon semiconductor detectors in degraded electron beams with a broad energy spectrum and a wide angular distribution. A comparison is made with diamond detector measurements, which is the dosimeter considered to give the best results provided that dose rate effects are corrected for. Two-dimensional relative absorbed dose distributions in electron beams (6-20 MeV) for intraoperative radiation therapy (IORT) are measured in a water phantom. To quantify deviations between the detectors, a dose comparison tool that simultaneously examines the dose difference and distance to agreement (DTA) is used to evaluate the results in low- and high-dose gradient regions, respectively. Uncertainties of the experimental measurement setup (+/- 1% and +/- 0.5 mm) are taken into account by calculating a composite distribution that fails this dose-difference and DTA acceptance limit. Thus, the resulting area of disagreement should be related to differences in detector performance. The dose distributions obtained with the diode are generally in very good agreement with diamond detector measurements. The buildup region and the dose falloff region show good agreement with increasing electron energy, while the region outside the radiation field close to the water surface shows an increased difference with energy. The small discrepancies in the composite distributions are due to several factors: (a) variation of the silicon-to-water collision stopping-power ratio with electron energy, (b) a more pronounced directional dependence for diodes than for diamonds, and (c) variation of the electron fluence perturbation correction factor with depth. For all investigated treatment cones and energies, the deviation is within dose-difference and DTA acceptance criteria of +/- 3% and +/- 1 mm, respectively. Therefore, p-type silicon diodes are well suited, in the sense that they give results in close agreement with diamond detectors, for practical measurements of relative absorbed dose distributions in degraded electron beams used for IORT.     

Lutzomyia longipalpis salivary gland homogenate impairs cytokine production and costimulatory molecule expression on human monocytes and dendritic cells

In this report, we describe an investigation of the effects of Lutzomyia longipalpis sand fly salivary gland homogenates (SGH) on cytokine production and expression of costimulatory molecules on human monocytes, macrophages (Ms), and dendritic cells (DCs). SGH of L. longipalpis induced an increase in interleukin-6 (IL-6), IL-8 and IL-12p40 production but a decrease in tumor necrosis factor alpha and IL-10 production by lipopolysaccharida (LPS)-stimulated monocytes. We also examined the expression of costimulatory molecules on the surface of monocytes, Ms, and DCs. Whereas SGH affected the expression of these molecules on monocytes and Ms, it had little effect on these molecules on DCs. However, when DCs were generated from human monocytes in the presence of SGH, SGH inhibited the expression of costimulatory molecules. In addition, a decrease in the maturation of DCs induced by CD40L was observed in the presence of SGH. Finally, preincubating SGH with human sera containing anti-SGH-specific antibodies abolished the effects of SGH on cytokine production by LPS-stimulated monocytes.     

Mapping of MRX81 in Xp11.2-Xq12 suggests the presence of a new gene involved in nonspecific X-linked mental retardation

X-linked nonspecific mental retardation (MRX) accounts for approximately 25% of mental retardation in males. A number of MRX loci have been mapped on the X chromosome, reflecting the complexity of gene action in central nervous system (CNS) specification and function. Eleven MRX genes have been identified, but many other causative loci remain to be refined to the single gene level. In 21 MRX families, the causative gene is located in the pericentromeric region; and we report here the identification by linkage analysis of a further such locus, MRX81. The new MRX locus was identified by two- and multi-point parametric analysis carried out on a large Italian family. Tight linkage of MRX81 to DNA markers ALAS2, DXS991, and DXS7132 was observed with a maximum LOD score of 3.43. Haplotype construction delineates an MRX81 critical region of 8 cM, the smallest MRX pericentromeric interval so far described, between DXS1039 and DXS1216, and placing it in Xp11.2-Xq12. So far, automated sequencing of two candidates in the region, the MRX gene oligophrenin (OPHN1) and the brain-specific ephrinB1 (EFNB1) gene, in DNA from affected males excluded their candidacy for MRX81, suggesting a novel disease gene.     

Spectral signature and heterodyne efficiency for different wavelengths in laser Doppler flowmetry

Laser Doppler perfusion monitoring and imaging technologies generate time traces and two-dimensional flow maps of the microcirculation. With the goal of reaching different tissue depths, these technologies are equipped with lassers operating at different wavelengths λ. The fact that the average scattering angle, at a single scattering event, between a photon and a red blood cell increases with λ is compensated for by a 1/λ effect in the scattering vector, rendering the average frequency shift virtually independent of the choice of wavelength. Monte Carlo simulations showed that the corresponding spectral signature of the Doppler signals for λ=632.8nm and 780nm were close to identical. The theoretical predictions were verified by calculating the centre-of-gravity (COG) frequency of the laser Doppler power spectral density for the two wavelengths from forearm and finger skin, representing a low and high perfusion area, respectively (forearm COG=123 against 121Hz, finger COG=220 against 212 Hz). When the wavelength changes from 632.8nm to 780nm, the heterodyne efficiency of the detector and, thereby, the inherent system amplifcation increase. For tissues with identical microvascular flow conditions, the output signal therfore tends to increase in magnitude when shifting to longer wavelengths.     

Neutrophilic granulocytes are the predominant cell type infiltrating pancreatic islets in contact with ABO-compatible blood

The poor outcome of intraportal islet transplantation may be explained by the instant blood-mediated inflammatory reaction (IBMIR), characterized by islet entrapment in blood clots, leucocyte infiltration and disruption of islet morphology. Here we employ a newly developed in vitro system to identify the blood cells involved in this process. Islets were mixed with ABO-compatible blood in heparinized tubes and incubated for various times up to 6 h. Clots were analysed immunohistochemically for detection of platelets (CD41a), leucocytes/lymphocytes (CD11b), granulocytes (CD16, lysozyme), neutrophilic granulocytes (neutrophil elastase), eosinophilic granulocytes (NaCN + H(2)O(2)), macrophages (CD68), dendritic cells (CD209/DC-SIGN), B cells (CD20) and T cells (CD4, CD8). Platelets were rapidly deposited around the islets in contact with the blood, reaching a maximum by 30 min. The first neutrophilic granulocytes appeared in the islets after 15 min, increased at 1 h and peaked at 2 h. Small numbers of macrophages were found infiltrating the islets already after 5 min, with a slight increase over time. However, control stainings of cultured islets and pancreas biopsies identified these cells as being largely of donor origin. No T cells, B cells, dendritic cells or eosinophilic granulocytes were detected during the 6 h observation time. Neutrophilic granulocytes were identified as the main infiltrating blood cell in islets exposed to blood, implying that these cells play a key role in clinical islet transplantation. Because islets are known to be exquisitely susceptible to oxidative stress, development of drugs targeting neutrophilic cytotoxicity could markedly improve the outcome of islet transplantation.     

Confounding factors in HLA-DW 2 typing of human leucocytes.

Three healthy HLA-B7 homozygous subjects were found with similar but not identical HLA-D antigens; one was DW 2 homozygous according to independent typing results. This could be an expression of "long" and "short" HLA-D antigens or be due to differences in weak antigens outside the HLA-D region. Two further healthy HLA-B7 homozygous subjects were studied; one was apparently heterozygous for DW 2, the other apparently carried no DW 2 antigen. Both could discriminate between different DW 2 homozygous test cells. Two such test cells--one from a patient with multiple sclerosis (MS) and the other from a man with two children with MS--gave variable and absurb reactions with cells from the two subjects in question. It is tentatively suggested that genes exist which, when present in both moities in a mixed leucocyte reaction (MLR), can impair the MLR and give false "typing" reactions. This might be more common among patients with MS and perhaps also some other diseases (certain arthritides, e.g. rheumatoid arthritis) than among healthy subjects and can complicate or make impossible the interpretation of HLA-D typing data. It could also explain the previously-described impaired MLR between cells from patients with these diseases.