Brain and Lung Imaging Correlation in Patients with COVID-19: Could the Severity of Lung Disease Reflect the Prevalence of Acute Abnormalities on Neuroimaging? A Global Multicenter Observational Study.

PURPOSE: Our aim was to study the association between abnormal findings on chest and brain imaging in patients with coronavirus disease 2019 (COVID-19) and neurologic symptoms. MATERIALS AND METHODS: In this retrospective, international multicenter study, we reviewed the electronic medical records and imaging of hospitalized patients with COVID-19 from March 3, 2020, to June 25, 2020. Our inclusion criteria were patients diagnosed with Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) infection with acute neurologic manifestations and available chest CT and brain imaging. The 5 lobes of the lungs were individually scored on a scale of 0-5 (0 corresponded to no involvement and 5 corresponded to >75% involvement). A CT lung severity score was determined as the sum of lung involvement, ranging from 0 (no involvement) to 25 (maximum involvement). RESULTS: A total of 135 patients met the inclusion criteria with 132 brain CT, 36 brain MR imaging, 7 MRA of the head and neck, and 135 chest CT studies. Compared with 86 (64%) patients without acute abnormal findings on neuroimaging, 49 (36%) patients with these findings had a significantly higher mean CT lung severity score (9.9 versus 5.8, P < .001). These patients were more likely to present with ischemic stroke (40 [82%] versus 11 [13%], P < .0001) and were more likely to have either ground-glass opacities or consolidation (46 [94%] versus 73 [84%], P = .01) in the lungs. A threshold of the CT lung severity score of >8 was found to be 74% sensitive and 65% specific for acute abnormal findings on neuroimaging. The neuroimaging hallmarks of these patients were acute ischemic infarct (28%), intracranial hemorrhage (10%) including microhemorrhages (19%), and leukoencephalopathy with and/or without restricted diffusion (11%). The predominant CT chest findings were peripheral ground-glass opacities with or without consolidation. CONCLUSIONS: The CT lung disease severity score may be predictive of acute abnormalities on neuroimaging in patients with COVID-19 with neurologic manifestations. This can be used as a predictive tool in patient management to improve clinical outcome.

Experimental Transmission of African Swine Fever (ASF) Low Virulent Isolate NH/P68 by Surviving Pigs.

African swine fever (ASF) has persisted in Eastern Europe since 2007, and two endemic zones have been identified in the central and southern parts of the Russian Federation. Moderate- to low-virulent ASF virus isolates are known to circulate in endemic ASF-affected regions. To improve our knowledge of virus transmission in animals recovered from ASF virus infection, an experimental in vivo study was carried out. Four domestic pigs were inoculated with the NH/P68 ASF virus, previously characterized to develop a chronic form of ASF. Two additional in-contact pigs were introduced at 72 days post-inoculation (dpi) in the same box for virus exposure. The inoculated pigs developed a mild form of the disease, and the virus was isolated from tissues in the inoculated pigs up to 99 dpi (pigs were euthanized at 36, 65, 99 and 134 dpi). In-contact pigs showed mild or no clinical signs, but did become seropositive, and a transient viraemia was detected at 28 days post-exposure (dpe), thereby confirming late virus transmission from the inoculated pigs. Virus transmission to in-contact pigs occurred at four weeks post-exposure, over three months after the primary infection. These results highlight the potential role of survivor pigs in disease maintenance and dissemination in areas where moderate- to low-virulent viruses may be circulating undetected. This study will help design better and more effective control programmes to fight against this disease.

Cytokine mRNA expression and pathological findings in pigs inoculated with African swine fever virus (E-70) deleted on A238L.

African swine fever virus (ASFV) induces a variety of immune responses and clinical forms in domestic pigs. As it is the only member of the Asfarviridae family, ASFV encodes many novel genes not encoded by other virus families. Among these genes, A238L may regulate the synthesis of pro-inflammatory cytokines, controlled mainly by NFkappaB and NFAT pathways. In this study, we inoculated two groups of pigs, one with the ASFV highly virulent E-70 isolate, deleted on A238L gene, and the other group with the parental E-70 isolate. No significant differences were observed in the clinical signs or pathology between both groups. However, the TNF-alpha mRNA expression was strongly enhanced in the PBMC from pigs inoculated with the virus deleted in A238L, reinforcing the role of the A238L gene in the inhibition of the NFkappaB pathway of expression of cytokines. No up-regulation of pro-inflammatory cytokines was observed in the PBMC of animals inoculated with the E-70 isolate, even though apoptosis and haemorrhages were evident and might be related to the presence of bystander monocyte-macrophages expressing these cytokines. Other studies using ASFV deleted in other genes inoculated in the natural hosts should be performed to gain further insight into the role of these genes in the pathogenesis of ASF.

Encephalitozoon microsporidia modulates p53-mediated apoptosis in infected cells.

Microsporidia are intracellular obligate parasites which have recently been found to be related to fungi. They have a unique extrusion apparatus that is able to inject the sporoplasm directly into the target cell without using receptors. Encephalitozoon microsporidia are a source of morbidity and mortality in humans. It has been suggested that microsporidia may modulate the host cell cycle and apoptosis. We report here that caspase-3 cleavage is inhibited at different times of Vero cell infection by Encephalitozoon microsporidia and that the phosphorylation and translocation of p53 to the nucleus, previous steps for the activation of this protein, do not occur after infection of Vero cells. Consequently, the transcriptional function of p53 is impaired during the infection cycle as demonstrated by luciferase reporter assays. Thus, to our knowledge, for the first time it is shown that an intracellular parasite may be able to multiply in the host cell without activating the p53 apoptotic pathway of that cell. However, changes in the expression of Bcl-2 or Bax levels were not observed.

Helical computed tomography angiography is the most efficient test to assess vascular calcifications in the iliac arterial sector in renal transplant candidates.

UNLABELLED: The increased scope of renal transplant indications has lead to a larger number of recipients with vascular problems due to arterial calcifications in the iliac region. Compared to magnetic resonance and conventional arteriography, helical computed tomography angiography (HCTA) accurately depicts arterial diseases, including the location and extent of arterial calcification. The objective of this study was to assess the value of HCTA with maximum-intensity-projection (MIP) reconstruction to evaluate iliac arterial calcifications and stenosis among candidates for renal transplantation. MATERIAL AND METHODS: From December 1997 to March 2002, 114 HCTA scans with MIP reconstruction were performed in candidates for renal transplantation. Included patients fulfilled some of the following conditions: (a) older than 55 years, (b) diabetic, (c) second transplants, and (d) obvious vascular calcifications on plain abdominal x-ray. RESULTS: Among the 114 patients, 33 (29%) were excluded for transplantation due to universal calcification of the iliac arterial sector, and 81 (71%) were included on the waiting list due to the presence of calcium-free areas for the vascular anastomosis. Transplantation, which was attempted in 28 of the 81 patients, was successful in 25 using the area programmed after HCTA analysis. The transplants failed in three cases because no calcium-free area could be found upon surgical examination. CONCLUSION: HCTA with MIP reconstruction makes it possible to draw an exact map of the arterial calcifications of the iliac arterial sector, allowing better recipient selection and accurate planning for the vascular anastomosis and placement of the renal graft.

Plaque assay for African swine fever virus on swine macrophages.

A plaque assay developed to detect the infection of African Swine Fever Virus on swine macrophages is described. Plaques were generated by all of the virus isolates tested. The method is suitable not only for virus titration but also for the selection of clones in protocols for isolation/purification of recombinant viruses.

African swine fever virus IAP homologue inhibits caspase activation and promotes cell survival in mammalian cells.

African swine fever virus (ASFV) A224L is a member of the inhibitor of apoptosis protein (IAP) family. We have investigated the antiapoptotic function of the viral IAP both in stably transfected cells and in ASFV-infected cells. A224L was able to substantially inhibit caspase activity and cell death induced by treatment with tumor necrosis factor alpha and cycloheximide or staurosporine when overexpressed in Vero cells by gene transfection. We have also observed that ASFV infection induces caspase activation and apoptosis in Vero cells. Furthermore, using a deletion mutant of ASFV lacking the A224L gene, we have shown that the viral IAP modulates the proteolytic processing of the effector cell death protease caspase-3 and the apoptosis which are induced in the infected cells. Our findings indicate that A224L interacts with the proteolytic fragment of caspase-3 and inhibits the activity of this protease during ASFV infection. These observations could indicate a conserved mechanism of action for ASFV IAP and other IAP family members to suppress apoptosis.

Influence of obesity on the appearance of proteinuria and renal insufficiency after unilateral nephrectomy.

BACKGROUND: Some patients develop proteinuria and progressive renal failure after unilateral nephrectomy, although the majority of patients maintain normal renal function. Reasons to explain this different evolution are not known. METHODS: A cross-sectional study was performed in 73 patients who had undergone unilateral nephrectomy 13.6 +/- 8.6 years before. Patients with morphologic abnormalities in the remaining kidney, systemic disorders, or abnormal renal function at the time of nephrectomy were excluded. All of the 73 included patients showed normal renal function and negative proteinuria at nephrectomy. The patient's medical records were reviewed, and clinical and analytical data throughout follow-up were obtained. RESULTS: Fifty-three out of the 73 patients (group I) showed a normal renal function and negative proteinuria at the cross-sectional study. The remaining 20 patients (group II) showed proteinuria (3.4 +/- 3.1 g/day). The time elapsed between nephrectomy and proteinuria appearance was 10.1 +/- 6.1 years. Thirteen patients of group II had developed renal insufficiency (serum creatinine at the cross-sectional study of 3.9 + 3.2 mg/dL) in addition to proteinuria. The time elapsed between proteinuria appearance and the onset of renal insufficiency was 4.1 +/- 4.3 years. Renal insufficiency showed a slowly progressive course in most of these patients. There were no significant differences between group I and group II patients in age, gender, renal function, or blood pressure at the time of nephrectomy. In contrast, group II patients showed a body mass index (BMI) that was significantly higher than group I at nephrectomy (31.6 +/- 5.6 vs. 24.3 +/- 3.7 kg/m(2), P < 0.001), at cross-sectional study (33.3 +/- 6.6 vs. 25.1 +/- 3.5 kg/m(2), P < 0.001), and throughout follow-up. Among the 14 obese (BMI > 30 kg/m(2)) patients at the time of nephrectomy, 13 (92%) developed proteinuria/renal insufficiency. In contrast, among the 59 patients with BMI < 30 kg/m(2), only 7 (12%) developed these complications (P < 0.001). Kaplan-Meier estimated probability of negative proteinuria and normal renal function 10 years after nephrectomy was 40 and 70%, respectively, in obese patients at nephrectomy. At 20 years after nephrectomy, these percentages were 8 and 35%, respectively. In contrast, in nonobese patients, the probability of negative proteinuria and normal renal function was 93 and 98%, respectively, at 10 years (P < 0.001) and 77 and 91%, respectively, at 20 years (P < 0.001). Multiple logistic regression analysis showed that the risk of developing renal disease was only statistically correlated with BMI at the time of unilateral nephrectomy (odds ratio 1.34, 1.03 to 1.76 CI). CONCLUSIONS: Obese patients are at risk for developing proteinuria and chronic renal failure after unilateral nephrectomy. Regular and long-term follow-up are recommended in these patients.

Mechanistic aspects of the induction of apoptosis by lauryl gallate in the murine B-cell lymphoma line Wehi 231.

The effect of lauryl gallate (antioxidant E-312) has been studied on the mouse B-cell lymphoma line Wehi 231. This compound is able to inhibit protein tyrosine kinases (PTKs) in whole cells and in crude extracts with a better efficiency than other well-known PTK inhibitors such as herbimycin or genistein. Initial events triggered upon the incubation of cells with lauryl gallate in phosphate-buffered saline (up to 1 h) include the inhibition of tyrosine phosphorylation, discharge of the mitochondrial transmembrane potential, and induction of mRNA for Bcl-2. Long-term cultures in complete medium supplemented with fetal calf serum (up to 24 h) in the presence of this compound exhibit clear apoptotic features such as increase in phosphatidylserine in the cell surface, decrease in the functionality of mitochondria, cytochrome c release to the cytosol, activation of caspases, hypodiploidy, and oligonucleosomal breakdown of DNA. Comparison between Wehi cells overexpressing Bcl-2 (Wehi-bcl-2) with Wehi-neo cells shows a delay in the manifestations of the apoptotic signs, indicating that Bcl-2 has a partial protective effect on the apoptosis induced by lauryl gallate. The proapoptotic effect of lauryl gallate is not dependent on DNA or protein synthesis, is not blocked by the chelation of calcium, and is not reverted by N-acetylcysteine.

Evanescent exostosis. A new case.

Much has been written about the natural history of osteochondromas, but there are only a few reports in the literature reflecting the spontaneous disappearance of this lesion. For that reason, we report an additional case which makes the total number of reported cases eleven, and also includes a review of the literature.

Inhibition of nuclear factor kappaB activation by a virus-encoded IkappaB-like protein.

Certain viruses have evolved mechanisms to counteract innate immunity, a host response in which nuclear factor kappaB (NF-kappaB) transcription factors play a central role. African swine fever virus encodes a protein of 28.2 kDa containing ankyrin repeats similar to those of cellular IkappaB proteins, which are inhibitors of NF-kappaB. Transfection of the African swine fever virus IkappaB gene inhibited tumor necrosis factor- or phorbol ester-induced activation of kappaB- but not AP-1-driven reporter genes. Moreover, African swine fever virus IkappaB co-immunoprecipitated with p65 NF-kappaB, and the purified recombinant protein prevented the binding of p65-p50 NF-kappaB proteins to their target sequences in the DNA. NF-kappaB activation induced by tumor necrosis factor, as detected by mobility shift assays or by transfection of kappaB-driven reporter genes, is impaired in African swine fever virus-infected cells. These results indicate that the African swine fever virus IkappaB gene homologue interferes with NF-kappaB activation, likely representing a new mechanism to evade the immune response during viral infection.

Vaccinia virus-induced apoptosis in immature B lymphocytes: role of cellular Bcl-2.

Apoptosis is a form of physiological cell death which can be initiated in response to various stimuli including virus infections. We show that vaccinia virus (VV) infection induces apoptosis in an immature B lymphocyte line, WEHI-231. In these cells, several VV-specific proteins were synthesized during the infection, but neither virus production nor viral DNA synthesis were detected. The intracellular levels of the proto-oncogene Bcl-2, which effectively protects cells from programmed cell death, were found to be down-regulated by the VV infection, suggesting that this down-regulation might be involved in the viral induction of apoptosis in WEHI-231 cells. Stable transfectants overexpressing human Bcl-2 were shown to be resistant to the apoptosis produced by the infection, a finding consistent with the proposed role for the down-regulation of endogenous Bcl-2 in VV-induced apoptotic death.

African swine fever virus-specific cytotoxic T lymphocytes recognize the 32 kDa immediate early protein (vp32).

African swine fever (ASF) virus-specific cytotoxic T lymphocyte (CTL) activity has been studied in a model in which SLA inbred minipigs were experimentally infected with an attenuated isolate of the virus. The CTL assays were performed using alveolar macrophages as target cells. The specific lysis is mediated by purified CD8+ lymphocytes but not by CD4+ cells and can be blocked by incubation with anti-SLA class I monoclonal antibodies. The purified CD8+ population produced high levels of interferon-gamma after ASF virus stimulation. In an attempt to define the viral proteins recognized by CTL, target cells infected with a recombinant vaccinia virus (VV) expressing the ASF virus p32, an immediate early protein during ASF virus replication, were recognized and lysed by CTL. This assay may be useful for VV recombinant screening in order to identify other potential target ASF virus proteins.

Inhibition of apoptosis by the African swine fever virus Bcl-2 homologue: role of the BH1 domain.

The function of the African swine fever virus (ASFV) bcl-2 homologue, gene A179L, in the regulation of apoptosis was investigated using as a model system the human myeloid leukemia cell line K562 induced to die by apoptosis with inhibitors of macromolecular synthesis, a process that is prevented by overexpression of human bcl-2. It is shown that transfection of K562 cells with the ASFV A179L gene protects these cells from apoptotic cell death induced by a combination of cycloheximide and actinomycin D or by treatment with cytosine arabinoside. To test the functional role of the highly conserved BH1 domain present in the A179L protein, the Gly residue at position 85 was mutated to Ala, since it has been shown that substitution of the corresponding Gly in human Bcl-2 abrogates its death-repressor activity. It was found that the Gly-to-Ala mutation in the BH1 domain of the viral protein abolished its capacity to protect the K562 cells from apoptosis, indicating that this Gly is essential for A179L action. This finding stresses the functional similarity of the BH1 domains of the viral protein and cellular Bcl-2.

Swine-reconstituted SCID mice as a model for African swine fever virus infection.

Injection of swine peripheral blood mononuclear cells into mice with severe combined immunodeficiency (SCID), resulted in the stable long-term establishment of a functional swine immune system (SCID-sw). Swine immunoglobulins were present in the serum of SCID-sw mice and swine cells were detected in the blood as well as in lymph nodes and spleen using monoclonal antibodies raised against cell subpopulations. Swine lymphocytes from reconstituted SCID mice responded in vitro to specific antigens or mitogens. When SCID-sw mice were challenged with African swine fever (ASF) virus, ASF virus-infected cells were detected in blood and spleen, and antiviral antibodies and virus-specific T cells were generated.

African swine fever virus guanylyltransferase.

The gene coding for the guanylyltransferase of African swine fever virus has been identified and sequenced. The gene, designated NP868R, is located within fragments EcoRI N' and D of the virus genome (BA71V strain) and encodes a protein with a predicted molecular mass of 99.9 kDa that shares significant similarity with the large subunit of both vaccinia and Shope fibroma virus capping enzymes, with percentages of identity of 20.6 and 21.8%, respectively. A protein of 95 kDa was induced in Escherichia coli cells transformed with a recombinant plasmid carrying the NP868R gene. The E. coli expressed protein, as well as a protein of the same molecular weight present in African swine fever virus particles, form a covalent complex with GTP that can be reversed by pyrophosphate, two characteristic reactions of guanylyltransferases. An examination of the amino acid sequences of the African swine fever virus, poxvirus, and yeast guanylyltransferases has revealed a common motif around a lysine residue at the amino-terminal part of the proteins [Y(V, A)X2K(T, A)DG] which resembles the adenylylation site of DNA ligases (Tomkinson, A. E., Totty, N. F., Ginsburg, M., and Lindahl, T. (1991). Proc. Natl. Acad. Sci. USA 88, 400-404). This lysine residue could be the guanylylation site in these enzymes.

Swine leukocyte antigen and macrophage marker expression on both African swine fever virus-infected and non-infected primary porcine macrophage cultures.

Swine leukocyte antigens (SLA) and a macrophage specific marker were monitored on porcine macrophages cultured with or without macrophage colony stimulatory factor (M-CSF) and on cells infected with African swine fever virus (ASFV). SLA expression was maximal either in the total cell extract or on the cell surface at 3-4 days of culture; after 4 days these values began to decrease. Fluorescence analyses of immunostained macrophages cultured with or without M-CSF indicated a major upward shift in the number of SLA Class I molecules on individual macrophages whereas for SLA Class II both a novel expression of Class II and an upward shift in the number of molecules per cell were evident. Infection of 3-day-old macrophage cultures with three different isolates of ASFV resulted in minor changes in surface expression of SLA Class I, SLA Class II, and macrophage markers. No differences in infection with ASFV was observed whether macrophages were SLA Class II positive or negative, nor was there blocking by anti-SLA Class I or Class II monoclonal antibodies of ASFV infection of cultured macrophages.

The structure of two IgMs showing different activity from a patient with Waldenstrom's macroglobulinaemia.

The two monoclonal IgMs (IgM1 and IgM2) were characterized from a patient Waldenström's macroglobulinaemia that resulted in a gammapathy. Heavy and light chains were isolated from the IgM. The complete primary structure of the two light chains and the NH2-terminal region of the two heavy chain molecules were determined. The sequence data indicated that the heavy and light chains from both IgMs belong to the same (III and II) lambda subgroups. By testing their antibody activity it was found by immunofluorescence and immunoblotting that only IgM2 reacts with an intermediate filament protein.

Interferon-gamma production by African swine fever virus-specific lymphocytes.

Peripheral blood mononuclear cells (PBMC) from inbred pigs that were immunized with autologous macrophages infected with the African swine fever (ASF) virus BA71V, a nonvirulent virus isolate, proliferated and produced interleukin-2 in response to homologous and heterologous isolates of the ASF virus. They produced, however, interferon (IFN) only when challenged in vitro with homologous or attenuated isolates of the ASF virus, but not with heterologous or virulent isolates. The IFN was pH 2 labile and was neutralized by specific serum to porcine recombinant IFN gamma.

Two IgM paraproteins bearing cryoglobulin and anti-smooth muscle activities in a patient with Waldenström's macroglobulinaemia.

A 52-year-old white male subject with typical clinical and laboratory findings of Waldenström's macroglobulinaemia is described. Two paraprotein peaks of IgM lambda class, with different physical and chemical properties and different amino acid compositions, in both heavy and light chains, were found in the patient's serum. One of the IgM components (M1) was a cryoglobulin, and the other (M2) showed strong antismooth muscle activity. As far as we know, this is the first report of double paraproteins each of which has different properties.