Cellular events of acute, resolving or progressive COVID-19 in SARS-CoV-2 infected non-human primates.

Understanding SARS-CoV-2 associated immune pathology is crucial to develop pan-effective vaccines and treatments. Here we investigate the immune events from the acute state up to four weeks post SARS-CoV-2 infection, in non-human primates (NHP) with heterogeneous pulmonary pathology. We show a robust migration of CD16 expressing monocytes to the lungs occurring during the acute phase, and we describe two subsets of interstitial macrophages (HLA-DR+CD206-): a transitional CD11c+CD16+ cell population directly associated with IL-6 levels in plasma, and a long-lasting CD11b+CD16+ cell population. Trafficking of monocytes is mediated by TARC (CCL17) and associates with viral load measured in bronchial brushes. We also describe associations between disease outcomes and high levels of cell infiltration in lungs including CD11b+CD16hi macrophages and CD11b+ neutrophils. Accumulation of macrophages is long-lasting and detectable even in animals with mild or no signs of disease. Interestingly, animals with anti-inflammatory responses including high IL-10:IL-6 and kynurenine to tryptophan ratios show less severe illness. Our results unravel cellular mechanisms of COVID-19 and suggest that NHP may be appropriate models to test immune therapies.

A Natural Alkaloid, ß-Carboline, as a One- and Two-Photon Responsive Fluorescent Photoremovable Protecting Group: Sequential Release of the Same or Different Carboxylic Acids.

The ß-carboline moiety, substituted at the C1 and C3 benzylic positions with a leaving group, has been demonstrated for the first time as a photoremovable protecting group for time-dependent sequential release of two (same or different) carboxylic acids upon one- and two-photon light irradiation. Density functional theory calculations suggest that the electronic environment of the ß-carboline moiety at C1 and C3 positions plays a key role in the rate of photorelease.

Observation of angle-dependent mode conversion and mode hopping in 2D annular antidot lattice.

We report spin-wave excitations in annular antidot lattice fabricated from 15 nm-thin Ni80Fe20 film. The nanodots of 170 nm diameters are embedded in the 350 nm (diameter) antidot lattice to form the annular antidot lattice, which is arranged in a square lattice with edge-to-edge separation of 120 nm. A strong anisotropy in the spin-wave modes are observed with the change in orientation angle (ϕ) of the in-plane bias magnetic field by using Time-resolved Magneto-optic Kerr microscope. A flattened four-fold rotational symmetry, mode hopping and mode conversion leading to mode quenching for three prominent spin-wave modes are observed in this lattice with the variation of the bias field orientation. Micromagnetic simulations enable us to successfully reproduce the measured evolution of frequencies with the orientation of bias magnetic field, as well as to identify the spatial profiles of the modes. The magnetostatic field analysis, suggest the existence of magnetostatic coupling between the dot and antidot in annular antidot sample. Further local excitations of some selective spin-wave modes using numerical simulations showed the anisotropic spin-wave propagation through the lattice.

All-inorganic quantum dot assisted enhanced charge extraction across the interfaces of bulk organo-halide perovskites for efficient and stable pin-hole free perovskite solar cells.

In spite of achieving high power conversion efficiency (PCE), organo-halide perovskites suffer from long term stability issues. Especially the grain boundaries of polycrystalline perovskite films are considered as giant trapping sites for photo-generated carriers and therefore play an important role in charge transportation dynamics. Surface engineering via grain boundary modification is the most promising way to resolve this issue. A unique antisolvent-cum-quantum dot (QD) assisted grain boundary modification approach has been employed for creating monolithically grained, pin-hole free perovskite films, wherein the choice of all-inorganic CsPbBr x I3-x (x = 1-2) QDs is significant. The grain boundary filling by QDs facilitates the formation of compact films with 1-2 µm perovskite grains as compared to 300-500 nm grains in the unmodified films. The solar cells fabricated by CsPbBr1.5I1.5 QD modification yield a PCE of ∼16.5% as compared to ∼13% for the unmodified devices. X-ray photoelectron spectral analyses reveal that the sharing of electrons between the PbI6 - framework in the bulk perovskite and Br- ions in CsPbBr1.5I1.5 QDs facilitates the charge transfer process while femtosecond transient absorption spectroscopy (fs-TAS) suggests quicker trap filling and enhanced charge carrier recombination lifetime. Considerable ambient stability up to ∼720 h with <20% PCE degradation firmly establishes the strategic QD modification of bulk perovskite films.

One- and Two-Photon Uncaging: Carbazole Fused o-Hydroxycinnamate Platform for Dual Release of Alcohols (Same or Different) with Real-Time Monitoring.

A one- and two-photon activated photoremovable protecting group (PRPG) was designed based on a carbazole fused o-hydroxycinnamate platform for the dual (same or different) release of alcohols. The mechanism for the dual release proceeds through a stepwise pathway and also monitors the first and second photorelease in real time by an increase in fluorescence intensity and color change, respectively. Further, its application in staining live neurons and ex vivo imaging with two-photon excitation is shown.

Loss of Smad4 in colorectal cancer induces resistance to 5-fluorouracil through activating Akt pathway.

BACKGROUND: Higher frequency of Smad4 inactivation or loss of expression is observed in metastasis of colorectal cancer (CRC) leading to unfavourable survival and contributes to chemoresistance. However, the molecular mechanism of how Smad4 regulates chemosensitivity of CRC is unknown. METHODS: We evaluated how the loss of Smad4 in CRC enhanced chemoresistance to 5-fluorouracil (5-FU) using two CRC cell lines in vitro and in vivo. Immunoblotting with cell and tumour lysates and immunohistochemical analyses with tissue microarray were performed. RESULTS: Knockdown or loss of Smad4 induced tumorigenicity, migration, invasion, angiogenesis, metastasis, and 5-FU resistance. Smad4 expression in mouse tumours regulated cell-cycle regulatory proteins leading to Rb phosphorylation. Loss of Smad4 activated Akt pathway that resulted in upregulation of anti-apoptotic proteins, Bcl-2 and Bcl-w, and Survivin. Suppression of phosphatidylinositol-3-kinase (PI3K)/Akt pathway by LY294002 restored chemosensitivity of Smad4-deficient cells to 5-FU. Vascular endothelial growth factor-induced angiogenesis in Smad4-deficient cells might also lead to chemoresistance. Low levels of Smad4 expression in CRC tissues correlated with higher levels of Bcl-2 and Bcl-w and with poor overall survival as observed in immunohistochemical staining of tissue microarrays. CONCLUSION: Loss of Smad4 in CRC patients induces resistance to 5-FU-based therapy through activation of Akt pathway and inhibitors of this pathway may sensitise these patients to 5-FU.

Diagnostic evaluation of supraclavicular lymphadenopathy.

Patients presented with the supraclavicular lymphadenopathy in the medicine department have a strong suspicion of serious illness like tuberculosis, sarcoidosis, toxoplasmosis and malignancy of lymphnode, blood, lung, upper GIT, breast, ovary, testes, and other sites of body. This prospective type of observational study carried out in the indoor and out patient department of medicine of Mymensingh Medical College Hospital over a period of 6 month from April 2011 to September 2011 to diagnose the causes of supraclavicular lymphadenopathy. Patient of either sex, 18 years or above presented with supraclavicular lymphadenopathy were included. Biopsy or FNAC were done. The study showed that mean age of the patient of supraclavicular lymphadenopathy that finally diagnosed as malignant was 49.7 years and that of non malignant was 33.7 years. Male patient have suffered more (60%) from malignant disease than that of female patient (40%). Discrete, hard, non tender either fixed or non fixed supraclavicular lymphadenopathy was found malignant (18 of 18 cases, 100%) and discrete, firm, tender lymphnode were found non malignant (5 of 5 cases, 100%). Increased frequency (11 of 28, 39.3%) of granulomatous inflammation from the tuberculoid lymphadenitis were found among the patient undergone supraclavicular lymphnode biopsy. FNAC result was also of simillar type and finally it was found that frequency of tuberculosis (20 of 53, 37.7%) was highest and bronchial carcima was the second most frequent diagnosis (14 of 53, 26.4%). This study showed that supraclavicular lymphadenopathy is associated mostly with serious disease like tuberculosis and malignancy.

Clinical profile of periodic paralysis.

This cross sectional descriptive study was done to find out common clinical presentations, etiologies and laboratory investigation abnormalities in patients of periodic paralysis. Study was carried out in 30 patients with an age range from 8 to 70 years who were enrolled from July 2008 to June 2009 in Mymensingh Medical College Hospital (MMCH) medicine unit. Individuals who were admitted with sudden onset generalized muscle weakness, had history of previous attack and serum potassium level <3mmol/l or >5.5mmol/l were included in this study. In this series, majority of the patients were male (66.67%). Male: female ratio was approximately 2:1. The mean age of the patients was 27.4±4.5 years. Majority (26.67%) of them were in age range of 31-40 years. About 30% of the patients experienced the first attack of paralysis at the age of 20-24 years. Majority of patients (53%) were from middle class family with occupation of private service (26.66%) and farmer (20%). Positive family history was reported in 20% of patients. Regarding the precipitating factors, majority of patients (83.3%) were related to high carbohydrate meal, 56.67% related to temperature, 41.67% to exercise. Flaccid muscle weakness with variables muscle power (MRC grade 4/5 to 2/5 in 60% and 1/5 to 0/5 in 40%) was found. Cerebellar functions, all modalities of sensations and functions of cranial nerves were intact in all patients. In this series, laboratory investigations revealed reduced serum potassium level (<3mmol/l) in 90% of patients. Serum potassium value >5.5mmol/l was found in only 3.33% of patients. Creatine kinase (MM) was raised in 23% of the patients and Thyroid stimulating hormone (TSH) level was 0.8-2mmol/l in 6% of the patients. More than half of the patients (56%) showed variable ECG changes. Impaired nerve conduction function was found in 28.00%. So, careful history taking, meticulous clinical examination and simple laboratory investigations is sufficient to make a prompt diagnosis and rapid management of patients with periodic paralysis.

Osteopetrosis.

A 15 years old Bangladeshi boy presented with hepatosplenomegaly, anaemia, multiple fractures (symptomatic and asymptomatic) without jaundice was investigated. Laboratory findings revealed leukoerythroblastic blood picture with reduced haemoglobin (7.7 gm/dl). Skeletal survey showed generalized increased bone density, sclerosed medulary space, Rugger-Jersey spine and diploic space filled with dense materials. Overlapping clinical features of both intermediate autosomal recessive and adult autosomal dominant variety of osteopetrosis were found in this patient but diagnosis were made on the basis of typical radiological finding which was mostly consistent with the adult autosomal dominant variety. The patient was treated conservatively and specialist consultation was taken in managing bony abnormalities. This patient was discharged with advised of subsequent follow-up.

Increased cell migration and plasticity in Nrf2-deficient cancer cell lines.

Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) expression is deregulated in many cancers. Genetic and biochemical approaches coupled with functional assays in cultured cells were used to explore the consequences of Nrf2 repression. Nrf2 suppression by Keap1-directed ubiquitylation or the expression of independent short hairpin RNA (shRNA)/siRNA sequences enhanced cellular levels of reactive oxygen species, Smad-dependent tumor cell motility and growth in soft agar. Loss of Nrf2 was accompanied by concomitant Smad linker region/C-terminus phosphorylation, induction of the E-cadherin transcriptional repressor Slug and suppression of the cell-cell adhesion protein E-cadherin. Ectopic expression of the wildtype but not dominant-negative Nrf2 suppressed the activity of a synthetic transforming growth factor-beta1-responsive CAGA-directed luciferase reporter. shRNA knock-down of Nrf2 enhanced the activity of the synthetic CAGA reporter, as well as the expression of the endogenous Smad target gene plasminogen activator inhibitor-1. Finally, we found that Nrf2/Smad3/Smad4 formed an immunoprecipitable nuclear complex. Thus, loss of Nrf2 increased R-Smad phosphorylation and R-Smad signaling, supporting the hypothesis that loss of Nrf2 in an oncogenic context-dependent manner can enhance cellular plasticity and motility, in part by using transforming growth factor-beta/Smad signaling.

The new high-temperature surface structure on reduced TiO2(001).

Scanning tunnelling microscopy, ultraviolet photoelectron spectroscopy and current imaging tunnelling spectroscopy (STM/UPS/CITS) were used to study the topographic and electronic structure of a high-temperature structure formed on the TiO(2)(001) surface after heating at 1173 K. The STM images revealed different domain-like ordering and periodicity on the surface in comparison to those observed previously. The UPS studies showed the presence of a surface state at energy about 1.1 eV below the Fermi level. This result was confirmed by the CITS data showing pronounced periodic maxima of the electron local density of states at energy around 1.1-1.2 eV below the Fermi level and located on top of every row of the new high-temperature structure. The CITS results recorded for small grains, which coexist with the observed structure, showed that their chemical composition is closer to the Ti(2)O(3) material than to TiO(2-x) for x << 1.

In vitro evaluation of UV opacity potential of Aloe vera L. gel from different germplasms.

In this study, lyophilized crude and methanolic extracts of aloe gel from different germplasms (S24, RM, TN, OR, and RJN) of Aloe vera L. were tested for their ultraviolet (UV) opacity potential. UV absorption profiles, sun protection factor (SPF), and percentage blocking of UVA and UVB were considered to test UV opacity potential. Both the extracts showed UV absorption and followed the same path in the wavelength range of 250-400 nm in all the germplasms. Methanolic extract showed a stronger absorptivity than the crude lyophilized extract. Among the tested germplasms, maximum UV opacity property with a SPF of 9.97% and 79.12% UVB blocking was obtained with RJN, whereas a poor response was evident in TN with a SPF of 1.37% and 28.5% UVB blocking at 4 mg/ml methanolic extract. To our knowledge the present work for the first time documents UV opacity properties of A. vera L. gel and opens up new vistas in Aloe gel characterization.

Smad7 induces hepatic metastasis in colorectal cancer.

Although Smad signalling is known to play a tumour suppressor role, it has been shown to play a prometastatic function also in breast cancer and melanoma metastasis to bone. In contrast, mutation or reduced level of Smad4 in colorectal cancer is directly correlated to poor survival and increased metastasis. However, the functional role of Smad signalling in metastasis of colorectal cancer has not been elucidated. We previously reported that overexpression of Smad7 in colon adenocarcinoma (FET) cells induces tumorigenicity by blocking TGF-beta-induced growth inhibition and apoptosis. Here, we have observed that abrogation of Smad signalling by Smad7 induces liver metastasis in a splenic injection model. Polymerase chain reaction with genomic DNA from liver metastases indicates that cells expressing Smad7 migrated to the liver. Increased expression of TGF-beta type II receptor in liver metastases is associated with phosphorylation and nuclear accumulation of Smad2. Immunohistochemical analyses have suggested poorly differentiated spindle cell morphology and higher cell proliferation in Smad7-induced liver metastases. Interestingly, we have observed increased expression and junctional staining of Claudin-1, Claudin-4 and E-cadherin in liver metastases. Therefore, this report demonstrates, for the first time, that blockade of TGF-beta/Smad pathway in colon cancer cells induces metastasis, thus supporting an important role of Smad signalling in inhibiting colon cancer metastasis.

HIV and complement: hijacking an immune defense.

The complement system is a central player of the innate immune system. Activation of the complement system protects the host against pathogens. However, uncontrolled synthesis can be detrimental to host. This concise review summarizes the current understanding of the mechanism(s) of complement activation, the mechanism of C3 regulation, and the role of complement in human immunodeficiency virus (HIV) pathogenesis with emphasis on the cross-talk between HIV and complement system in NeuroAIDS and HIV-associated nephropathy (HIVAN).

Restoration of TGF-beta signalling reduces tumorigenicity in human lung cancer cells.

Members of the transforming growth factor-beta (TGF-beta) family regulate a wide range of biological processes including cell proliferation, migration, differentiation, apoptosis, and extracellular matrix deposition. Resistance to TGF-beta-mediated tumour suppressor function in human lung cancer may occur through the loss of type II receptor (TbetaRII) expression. In this study, we investigated the expression pattern of TbetaRII in human lung cancer tissues by RT-PCR and Western blot analyses. We observed downregulation of TbetaRII in 30 out of 46 NSCLC samples (65%) by semiquantitative RT-PCR. Western blot analyses with tumour lysates showed reduced expression of TbetaRII in 77% cases. We also determined the effect of TbetaRII expression in lung adenocarcinoma cell line (VMRC-LCD) that is not responsive to TGF-beta due to lack of TbetaRII expression. Stable expression of TbetaRII in these cells restored TGF-beta-mediated effects including Smad2/3 and Smad4 complex formation, TGF-beta-responsive reporter gene activation, inhibition of cell proliferation and increased apoptosis. Clones expressing TbetaRII showed reduced colony formation in soft-agarose assay and significantly reduced tumorigenicity in athymic nude mice. Therefore, these results suggest that reestablishment of TGF-beta signalling in TbetaRII null cells by stable expression of TbetaRII can reverse malignant behaviour of cells and loss of TbetaRII expression may be involved in lung tumour progression.

Over-expression of cyclin D1 regulates Cdk4 protein synthesis.

Increased Cdk4 expression occurs coincident with over-expression of cyclin D1 in many human tumours and tumourigenic mouse models. Here, we investigate both in vivo and in vitro the mechanism by which Cdk4 expression is regulated in the context of cyclin D1 over-expression. Cdk4 mRNA levels in cyclin D1-over-expressing tissue and cultured cells were unchanged compared with controls. In contrast, Cdk4 protein levels were increased in cyclin D1-over-expressing tissue and cells versus their respective controls. This increase was not due to altered protein stability, but appeared to be due to an increase in Cdk4 protein synthesis. We also performed immunoprecipitation and in vitro kinase assays to demonstrate an increase in cyclin D1-Cdk4 complex formation and associated kinase activity. Blocking cyclin D1 expression resulted in diminished Cdk4 protein but not mRNA levels. These findings suggest a mechanism by which Cdk4 expression is increased in the context of cyclin D1 over-expression during tumourigenesis.

Acid-catalyzed cyclization of 2,3-dibenzylidenesuccinates: synthesis of lignans (+/-)-cagayanin and (+/-)-galbulin.

Acid-catalyzed cyclizations of E,E-dibenzylidenesuccinate esters have been developed as an efficient synthetic route to 1-aryl-1,2-dihydronaphthalenes. This reaction has been used in the synthesis of the naturally occurring lignans (+/-)-cagayanin and (+/-)-galbulin.

Oncogenic ras represses transforming growth factor-beta /Smad signaling by degrading tumor suppressor Smad4.

The loss of growth-inhibitory responses to transforming growth factor-beta (TGF-beta) is a frequent consequence of malignant transformation. Smad2, Smad3, and Smad4 proteins are important mediators of the antiproliferative responses to TGF-beta and may become inactivated in some human cancers. Epithelial cells harboring oncogenic Ras mutations often exhibit a loss of TGF-beta antiproliferative responses. To further investigate the effect of oncogenic Ras in TGF-beta signaling, we used an isopropyl-1-thio-beta-d-galactopyranoside-inducible expression system to express Ha-Ras(Val-12) in intestinal epithelial cells. Induction of Ha-Ras(Val-12) caused a decrease in the level of Smad4 expression, inhibited TGF-beta-induced complex formation between Smad2/Smad3 and Smad4, blocked Smad4 nuclear translocation, inhibited the TGF-beta-mediated decrease in [(3)H]thymidine incorporation, and repressed TGF-beta-activated transcriptional responses. The withdrawal of isopropyl-1-thio-beta-d-galactopyranoside or the addition of an inhibitor of the ubiquitin-proteasome pathway restored the Smad4 level and TGF-beta-induced Smad complex formation. Forced expression of Smad4 resulted in partial recovery of the TGF-beta-mediated growth inhibition and transcriptional responses in the presence of oncogenic Ras. Further, PD98059, a specific inhibitor of the MEK/ERK/mitogen-activated protein kinase pathway prevented the Ras-induced decrease in Smad4 expression and complex formation. Our results suggest a novel mechanism by which oncogenic Ras represses TGF-beta signaling by mitogen-activated protein kinase-dependent down-regulation of Smad4, thereby subverting the tumor suppressor function of TGF-beta.

Induction of heme oxygenase 1 in radiation nephropathy: role of angiotensin II.

Datta, P. K., Moulder, J. E., Fish, B. L., Cohen, E. P. and Lianos, E. A. Induction of Heme Oxygenase 1 in Radiation Nephropathy: Role of Angiotensin II. Radiat. Res. 155, 734-739 (2001). In a rat model of radiation-induced nephropathy, we investigated changes in expression of heme oxygenase 1 (Hmox1, also known as HO-1), an enzyme that catalyzes conversion of heme into biliverdin, carbon monoxide and iron. The study explored whether radiation induces Hmox1 expression in the irradiated kidney and whether angiotensin II (AII) mediates Hmox1 expression in glomeruli isolated from irradiated kidneys. To assess the effects of radiation on Hmox1 expression, rats received 20 Gy bilateral renal irradiation and were randomized to groups receiving an AII type 1 (AT(1)) receptor antagonist (L-158,809) or no treatment. Drug treatment began 9 days prior to bilateral renal irradiation and continued for the duration of the study. Estimation of Hmox1 levels in glomerular protein lysates assessed by Western blot analysis revealed a significant increase in Hmox1 protein at 50 and 65 days postirradiation. In animals treated with the AT(1) receptor antagonist, there was no induction of Hmox1, suggesting that AII may be a mediator of Hmox1 induction. To confirm that AII stimulates Hmox1 expression, animals were infused with 200, 400 or 800 ng/kg min(-1) of AII for 18-19 days, and Hmox1 protein levels in glomeruli were assessed. There was a significant induction of Hmox1 in glomeruli of animals infused with 800 ng/kg min(-1) of AII. These studies demonstrate that glomerular Hmox1 expression is elevated in the middle phase of radiation nephropathy and that AII can increase glomerular Hmox1 levels.