TLR5-mediated sensing of gut microbiota is necessary for antibody responses to seasonal influenza vaccination.

Systems biological analysis of immunity to the trivalent inactivated influenza vaccine (TIV) in humans revealed a correlation between early expression of TLR5 and the magnitude of the antibody response. Vaccination of Trl5(-/-) mice resulted in reduced antibody titers and lower frequencies of plasma cells, demonstrating a role for TLR5 in immunity to TIV. This was due to a failure to sense host microbiota. Thus, antibody responses in germ-free or antibiotic-treated mice were impaired, but restored by oral reconstitution with a flagellated, but not aflagellated, strain of E. coli. TLR5-mediated sensing of flagellin promoted plasma cell differentiation directly and by stimulating lymph node macrophages to produce plasma cell growth factors. Finally, TLR5-mediated sensing of the microbiota also impacted antibody responses to the inactivated polio vaccine, but not to adjuvanted vaccines or the live-attenuated yellow fever vaccine. These results reveal an unappreciated role for gut microbiota in promoting immunity to vaccination.

Acoustic shock waves emitted from two interacting laser generated plasmas in air.

We present an acoustic detection technique to study the interaction of two shock waves emitted by two nearby, simultaneous, laser-induced air-breakdown events that resembles the phenomenon of interaction of fluids. A microphone is employed to detect the acoustic shock wave (ASW) from the interaction zone. The experiments were done at various separation distances between the two plasma sources. The incident laser energy of the sources is varied from 25 to 100 mJ in ratios from 1:1 to 1:4. The peak sound pressure of the ASW was compared between the single and dual plasma sources, showing that the pressures are higher for the dual plasma source than that of the single plasma. The evolution of peak sound pressures is observed to depend on (a) the pulse energy of the sources and (b) the plasma separation distance, d. For the equal energy sources, the peak sound pressures increased linearly up to a certain plasma separation distance d, beyond which the pressures saturated and decayed. For the case of unequal energy sources, the peak sound pressures showed an interesting response of increase, saturation, decay, and further increase with plasma separation distance d. These observations indicate the dynamics of acoustic wave interactions across the interaction zone of the two sources depend on the input laser pulse energy as well as the plasma separation distance d.

Nitrate and Nitrite Reductions at Copper(II) Sites: Role of Noncovalent Interactions from Second-Coordination-Sphere.

Reductions of nitrate and nitrite (NOx-) are of prime importance in combatting water pollution arising from the excessive use of N-rich fertilizers. While examples of NOx- reductions are known, this report illustrates hydrazine (N2H4)-mediated transformations of NOx- to nitric oxide (NO)/nitrous oxide (N2O). For nitrate reduction to NO, initial coordination of the weakly coordinating NO3- anion at [(mC)CuII]2+ cryptate has been demonstrated to play a crucial role. A set of complementary analyses (X-ray diffraction and Fourier-transform infrared spectroscopy (FTIR), UV-vis, and NMR spectroscopies) on NO3--bound metal-cryptates [(mC)MII(NO3)](ClO4) (1-M, M = Cu/Zn) demonstrates the binding of NO3- through noncovalent (NH···O, CH···O, and anion···π) and metal-ligand coordinate interactions. Subsequently, reactions of [(mC)CuII(14/15NO3)](ClO4) (1-Cu or 1-Cu/15N) with N2H4·H2O have been illustrated to reduce 14/15NO3- to 14/15NO. Intriguingly, in the absence of the second-coordination-sphere interactions, a closely related coordination motif [(Bz3Tren)CuII]2+ (in 3-Cu) does not bind NO3- and is unable to assist in N2H4·H2O-mediated NO3- reduction. In contrast, nitrite coordinates at the tripodal CuII sites in both [(mC)CuII]2+ and [(Bz3Tren)CuII]2+ irrespective of the additional noncovalent interactions, and hence, the N2H4 reactions of the copper(II)-nitrite complexes [(mC)CuII(O14/15NO)]+ and [(Bz3Tren)CuII(O14/15NO)]+ (in 2-Cu/4-Cu) result in a mixture of 14/15NO and N14/15NO.

Infections in Infants with SCID: Isolation, Infection Screening, and Prophylaxis in PIDTC Centers.

PURPOSE: The Primary Immune Deficiency Treatment Consortium (PIDTC) enrolled children with severe combined immunodeficiency (SCID) in a prospective natural history study of hematopoietic stem cell transplant (HSCT) outcomes over the last decade. Despite newborn screening (NBS) for SCID, infections occurred prior to HSCT. This study's objectives were to define the types and timing of infection prior to HSCT in patients diagnosed via NBS or by family history (FH) and to understand the breadth of strategies employed at PIDTC centers for infection prevention. METHODS: We analyzed retrospective data on infections and pre-transplant management in patients with SCID diagnosed by NBS and/or FH and treated with HSCT between 2010 and 2014. PIDTC centers were surveyed in 2018 to understand their practices and protocols for pre-HSCT management. RESULTS: Infections were more common in patients diagnosed via NBS (55%) versus those diagnosed via FH (19%) (p = 0.012). Outpatient versus inpatient management did not impact infections (47% vs 35%, respectively; p = 0.423). There was no consensus among PIDTC survey respondents as to the best setting (inpatient vs outpatient) for pre-HSCT management. While isolation practices varied, immunoglobulin replacement and antimicrobial prophylaxis were more uniformly implemented. CONCLUSION: Infants with SCID diagnosed due to FH had lower rates of infection and proceeded to HSCT more quickly than did those diagnosed via NBS. Pre-HSCT management practices were highly variable between centers, although uses of prophylaxis and immunoglobulin support were more consistent. This study demonstrates a critical need for development of evidence-based guidelines for the pre-HSCT management of infants with SCID following an abnormal NBS. TRIAL REGISTRATION: NCT01186913.

Fingolimod retains cytolytic T cells and limits T follicular helper cell infection in lymphoid sites of SIV persistence.

Lymph nodes (LN) and their resident T follicular helper CD4+ T cells (Tfh) are a critical site for HIV replication and persistence. Therefore, optimizing antiviral activity in lymphoid tissues will be needed to reduce or eliminate the HIV reservoir. In this study, we retained effector immune cells in LN of cART-suppressed, SIV-infected rhesus macaques by treatment with the lysophospholipid sphingosine-1 phosphate receptor modulator FTY720 (fingolimod). FTY720 was remarkably effective in reducing circulating CD4+ and CD8+ T cells, including those with cytolytic potential, and in increasing the number of these T cells retained in LN, as determined directly in situ by histocytometry and immunohistochemistry. The FTY720-induced inhibition of T cell egress from LN resulted in a measurable decrease of SIV-DNA content in blood as well as in LN Tfh cells in most treated animals. In conclusion, FTY720 administration has the potential to limit viral persistence, including in the critical Tfh cellular reservoir. These findings provide rationale for strategies designed to retain antiviral T cells in lymphoid tissues to target HIV remission.

Effect of focusing element-induced aberrations on filamentation and supercontinuum emission in ambient air.

Femtosecond laser pulse induced filamentation in atmosphere is susceptible to a number of input laser, focusing optics and medium characteristics. Filamentation of fs pulses in atmosphere is an intense propagation regime where the focusing geometry used to focus the fs laser pulses play an important role influencing the filament intensity and the associated supercontinuum. We identified different optical elements used for focusing the fs laser pulses leading to filamentation in air and classified them according to the induced aberrations. To clearly identify the role of aberrations, all the optical elements were taken to have same focal length. The subsequent filament structure and emissions from the filament were correlated with the aberrations induced by optical element revealed stark differences. The onset of the filamentation, its longitudinal intensity and the associated supercontinuum emission (SCE) have varied drastically with the aberrations induced by the focusing optics. A systematic study directed to choose and identify suitable optical elements according to the usage of the fs pulses for a specific filamentation regime is presented.

Spatio-temporal characterization of ablative Cu plasma produced by femtosecond filaments.

We present the spatial and temporal characterization of the copper (Cu) plasma produced by the femtosecond laser filaments. The filaments of various lengths and intensities were generated with the aid of three different focusing lenses. Further, the filamentation induced breakdown spectroscopy (FIBS) measurements were carried out for each filament at three different positions along the length of the filament. The filaments were spatially characterized by estimating the plasma temperature and electron density. Our investigation has demonstrated that the centre of the filament is the best to obtain a maximum signal. Both the spectral line intensity and their persistence time are highest for the center of the filament. The enhanced persistence and the scalability of the spectral line intensity tested across different focusing geometries can boost the application of this technique in various fields.

Supercontinuum generation from zinc borate glasses: bandgap versus rare-earth doping.

We report an enhanced supercontinuum generation (SCG) from a rare-earth (Pr3+) doped low bandgap zinc borate glass when excited with 60 fs pulses from a Ti-sapphire laser. The emission associated with the absorption bands due to Pr3+ doping (around 420-500 and 580-600 nm) is observed to assist the enhanced SCG.

Input pulse duration effect on laser-induced underwater acoustic signals.

The characteristics of laser-induced underwater acoustic signals (UASs) generated by focused 10 ns and 30 ps laser pulses of different energies under similar experimental conditions are compared. Time domain signals and time-frequency analysis of the UASs were used to understand the role of input pulse duration and energy on the evolution of UAS. In the time domain, the peak-to-peak (${{\rm P}_k} {-} {{\rm P}_k}$) overpressure of the UAS decreases, and the arrival time (${{\rm A}_t}$) increases as a function of propagation distance for both ns and ps laser-induced breakdown (LIB) of water. With increasing incident energy of both ns and ps laser pulses, the ${{\rm P}_k} {-} {{\rm P}_k}$ overpressures of acoustic signals increase almost linearly. In the time-frequency domain, the spectrogram obtained via short-time Fourier transform provides spectral information and ${{\rm A}_t}$ of both direct and reflected signals simultaneously. The spectrogram revealed that the transient UAS has broad acoustic spectra spanning from 10 to 800 kHz, perpendicular to the laser propagation direction. The initial acoustic impulse resulted in two major frequencies centered around 105 and 690 kHz with a standard error of 30 kHz. Upon reflection from the water-air interface, only the peak frequency corresponding to ${\sim}{105}\;{\rm kHz}$ was reflected, while the longer frequency was observed to dissipate. Our results demonstrate that the ns-LIB is more suitable for applications compared to the ps-LIB owing to stronger acoustic impulse of both direct and reflected signals.

Green synthesis of silver, iron and gold nanoparticles of lycopene extracted from tomato: their characterization and cytotoxicity against COLO320DM, HT29 and Hella cell.

Our study aimed at development of Silver, Iron and Gold nanoparticles of Lycopene isolated from tomato by using green synthesis technique and to evaluate its anticancer potential against colorectal and cervical cancer. Lycopene was extracted by benzene extraction method and the silver, iron and gold nanoparticles were developed by green synthesis method. 1% aqueous extract of isolated Lycopene was mixed with 1% solutions of AgNO3, FeCl3 and HAuCl4 solutions and incubated at ambient temperature for 3-4 h separately and observed for the color change which is an indicative of formation of the nanoparticles. The prepared nanoparticles were characterized by FTIR, SEM, XRD analysis and evaluated for their antimicrobial potential. The cytotoxicity studies were carried out by in vitro assay like MTT, SRB and Tryphan blue method against Colo 320 DM, HT 29, and Hella. SEM showed nanosized particles of 50-100 nm range, whereas no antimicrobial activity was exhibited by the prepared nanoparticles. In MTT assay the LyAgNP showed maximum 41.41 ± 0.4124% inhibition against COLO320DM, whereas LyGNP exhibited 41.47 ± 0.4469% inhibition against HT 29 and LyAgNP showed 40.9 ± 0.6908% inhibition against Hella cells. In SRB assay LyAgNP showed maximum 82.68 ± 1.1798% inhibition against COLO320DM, whereas LyGNP exhibited maximum 91.21 ± 0.2372% inhibition against HT29 and 87.98 ± 0.5878% inhibition against Hella cells. In tryphan blue assay against COLO320DM, HT29 and Hella cells, the maximum inhibition exhibited by the prepared nanoparticles were observed as LyGNP 83.45 ± 0.4694%, LyAgNP 88.05 ± 0.1870% and LyAgNP65.47 ± 0.4766%. We conclude that the developed nanoparticles of Lycopene exhibited potential anticancer activity against Colorectal and cervical cancer cell as compared with pure Lycopene.

Safety of Baricitinib 4 mg for the Treatment of Moderate to Severe Rheumatoid Arthritis.

OBJECTIVE: We evaluated the safety of baricitinib 4 mg at 24 weeks for the treatment of moderate to severe rheumatoid arthritis (RA). METHODS: Multiple databases were searched from inception up to November 26, 2019 for randomized controlled trials comparing baricitinib 4 mg with placebo for the treatment of moderate to severe RA. The safety outcomes of interest were the incidence of serious adverse events, adverse events leading to study discontinuation, all infections, and serious infections. Adjusted risk ratios (RRs) with 95% confidence intervals (CIs) were pooled for safety outcomes. The Cochrane tool was used to assess the risk of bias. RESULTS: This analysis included four randomized controlled trials with 3106 patients. For serious adverse events, the pooled RR (95% CI) was 1.09 (0.76-1.57). For adverse events leading to study discontinuation, the pooled RR (95% CI) was 1.41 (0.94-2.11). For all reported infections, the pooled RR (95% CI) was 1.24 (1.10-1.40), For serious infections, pooled RR (95% CI) was 0.97 (0.51-2.57). CONCLUSIONS: Patients with RA taking 4 mg baricitinib daily did have an increased risk of infections; however, the incidence of serious adverse events, adverse events leading to study discontinuation, or serious infections were not significantly different in patients treated with baricitinib 4 mg compared with placebo.

Colon targeted dosage form of Capecitabine using folic acid anchored modified carbon nanotube: in vitro cytotoxicity, apoptosis and in vivo roentgenographic study.

OBJECTIVE: Development of dosage form comprising of Capecitabine loaded carbon nanotubes for its targeted delivery to the colon. METHOD: Single walled carbon nanotubes (SWCNT) were functionalized by -COOH and Chitosan along with Folic acid. Capecitabine was loaded in these SWCNT's, and the system was analyzed by FTIR, SEM and Raman spectroscopy. Percent drug loading was assessed and the cytotoxicity (COLO320DM and HT29) was verified by using MTT and SRB assay. The apoptosis study was carried out by flowcytometry. The system was enclosed in an enteric coated capsule with pH sensitive polymers and characterized for invitro disintegration, dissolution and invivo roentgenographic studies. RESULTS: FTIR, Raman and XRD studies indicated the confirmation of attachments, whereas SEM exhibited size range of 200-500 nm. Drug loading capacity was observed to be 94.63 ± 1.07%. Cytotoxicity studies of Capecitabine and FA-CHI-F-SWCNT-Capecitabine against COLO320DM by using MTT assay showed that FA-CHI-F-SWCNT- Capecitabine exhibited 86.45 ± 0.5788% inhibition whereas pure Capecitabine showed 50.52 ± 0.3106% inhibition. Against HT29, the % inhibition was observed to be 82.76 ± 0.4668% and 56.41 ± 0.2316% respectively for FA-CHI-F-SWCNT-Capecitabine and pure Capecitabine. In case of SRB assay of COLO320DM, the FA-CHI-F-SWCNT-Capecitabine exhibited 89.62 ± 0.4095% inhibition and Capecitabine showed 84.36 ± 0.2559% inhibition, whereas against HT29, FA-CHI-F-SWCNT-Capecitabine showed 81.36 ± 0.2958% inhibition and Capecitabine exhibited 90.62 ± 0.4196% inhibition. CONCLUSION: FA-CHI-F-SWCNT loaded system revealed better cytotoxicity as compared with pure Capecitabine against two different cell lines. Invivo studies revealed that the prepared capsule formulation remained intact in the stomach thereby preventing drug release in the gastric milieu.

Phenol Reduces Nitrite to NO at Copper(II): Role of a Proton-Responsive Outer Coordination Sphere in Phenol Oxidation.

In the view of physiological significance, the transition-metal-mediated routes for nitrite (NO2-) to nitric oxide (NO) conversion and phenol oxidation are of prime importance. Probing the reactivity of substituted phenols toward the nitritocopper(II) cryptate complex [mC]Cu(κ2-O2N)(ClO4) (1a), this report illustrates NO release from nitrite at copper(II) following a proton-coupled electron transfer (PCET) pathway. Moreover, a different protonated state of 1a with a proton hosted in the outer coordination sphere, [mCH]Cu(κ2-O2N)(ClO4)2 (3), also reacts with substituted phenols via primary electron transfer from the phenol. Intriguingly, the alternative mechanism operative because of the presence of a proton at the remote site in 3 facilitates an unusual anaerobic pathway for phenol nitration.

NAPDH Oxidase-Specific Flow Cytometry Allows for Rapid Genetic Triage and Classification of Novel Variants in Chronic Granulomatous Disease.

PURPOSE: Chronic granulomatous disease (CGD) is an innate immune deficiency, primarily affecting the phagocytic compartment, and presenting with a diverse phenotypic spectrum ranging from severe childhood infections to monogenic inflammatory bowel disease. Dihydrorhodamine (DHR) flow cytometry is the standard diagnostic test for CGD, and correlates with NADPH oxidase activity. While there may be genotype correlation with the DHR flow pattern in some patients, in several others, there is no correlation. In such patients, assessment by flow cytometric evaluation of NADPH oxidase-specific (NOX) proteins provides a convenient and rapid means of genetic triage, though immunoblotting has long been used for this purpose. METHODS AND RESULTS: We describe the clinical utility of the NOX flow cytometry assay through assessment of X-linked and autosomal recessive CGD patients and their first-degree relatives. The assessment of specific NOX proteins was correlated with overall NADPH oxidase function (DHR flow), clinical phenotype and genotype. NOX-specific protein assessment is a valuable adjunct to DHR assessment and genotyping to classify and characterize CGD patients. CONCLUSIONS: The atypical clinical presentation of some CGD patients can make genotype-phenotype correlation with DHR flow data challenging. Genetic testing, while useful for confirmation of diagnosis, can take several weeks, and in some patients does not provide a conclusive answer. However, NADPH-oxidase-specific protein flow assessment offers a rapid alternative to identification of the underlying genetic defect in cellular subsets, and can be utilized as a reflex test to an abnormal DHR flow. Further, it can provide insight into correlation between oxidative burst relative to protein expression in granulocytes and monocytes.

Pathological, ultrasonographic, and electrophysiological characterization of clinically diagnosed cases of pure neuritic leprosy.

A subset of neuritic form of leprosy, called pure neuritic leprosy (PNL), seen in a minority of leprosy patients, is characterized by peripheral neuropathy without skin lesions and an absence of acid-fast bacilli on skin smears. Patients with PNL are often started on drug therapy without confirmation of diagnosis. We, therefore, did a prospective study of clinically diagnosed PNL patients with correlation of ultrasonographic and biopsy findings. A total of 100 consecutive patients with PNL, diagnosed according to the consensus case definition, were included in the study. All patients underwent nerve conduction study, peripheral nerve ultrasonography, and sural nerve biopsy. Multiple mononeuropathies were present in 75% of cases, mononeuropathy in 18%, and polyneuropathy in the remaining 7%. Compared to clinical examination, ultrasonographic assessment of the peripheral nerves was not only better at the detection of thickening but also helped in characterization of their fascicular architecture, echogenicity, and vascularity. A total of 32 cases were confirmed on nerve biopsy, out of which 75% had demonstrable lepra bacilli. Cranial nerve involvement, presence of trophic ulcers, and bilateral thickening of the great auricular nerve were significantly associated with the positivity of lepra bacilli. A significant improvement in the disability score happened after multidrug therapy. A comprehensive electrophysiologic, ultrasonographic, and histological evaluation may be helpful in establishing a diagnosis of PNL with greater confidence, while ruling out other non-leprosy diagnoses.

Development of a stable chemically cross-linked erythropoietin dimer for use in the quality control of erythropoietin therapeutic products.

Erythropoietin (EPO) is a glycoprotein hormone which promotes red cell replenishment and is also a global biotherapeutic medicine widely used to treat anaemia resulting, for example, from chemotherapy. Requirements of the European Pharmacopoeia stipulate that the level of dimer must be quantified in clinical EPO products (with a limit of 2%). Quantification is hampered by the lack of reference preparations containing stable measurable levels of EPO dimer, but the reproducible generation of a stable dimerised EPO preparation is challenging. We describe here the development of a lyophilised, chemically cross-linked EPO preparation, which has good stability and may be used for calibration and system suitability assurance for the size exclusion chromatographic separation of EPO preparations. Graphical abstract.

Gangrenous Necrotizing Cutaneous Mucormycosis in an Immunocompetent Neonate: A Case Report from Oman.

We describe a case of progressive cutaneous mucormycosis, which caused gangrenous necrotizing fasciitis, in an immunocompetent neonate from Oman. Extensive wound contamination and a favorable environment for the growth of Mucorales were the predisposing factors. Early aggressive management including frequent wound debridement, targeted pharmacotherapy with liposomal amphotericin B and supportive care is pivotal for improved outcomes.

Etiologic spectrum and prognosis in noncompressive acute transverse myelopathies: An experience of 80 patients at a tertiary care facility.

INTRODUCTION: We evaluated the spectrum of acquired demyelinating and inflammatory disorders in patients presenting with an acute transverse myelopathy. We also studied differences between an acute idiopathic transverse myelitis and myelitis resulting from other etiologies. MATERIALS AND METHODS: Eighty consecutive patients with acute transverse myelopathy were included. At inclusion, clinical profile, serum and cerebrospinal fluid parameters, brain and spinal cord magnetic resonance imaging, and visual evoked potentials were obtained. All patients were given methylprednisolone therapy. Patients were followed up for 6 months. Outcome was assessed using modified Barthel index. A modified Barthel index score of ≤12 indicated a poor prognosis. RESULTS: Majority (n = 49; 61.25%) of patients had idiopathic acute transverse myelitis. Eleven cases had neuromyelitis optica spectrum disorders (8 had anti-aquaporin antibody positivity). Multiple sclerosis was diagnosed in 7 cases. Eight cases had infectious or parainfectious myelitis. Longitudinally extensive transverse myelitis was noted in 66 (82.5%) patients. Seventeen patients had abnormalities in the brain. Majority of patients improved following methylprednisolone therapy. On univariate analysis, delay in administering methylprednisolone therapy, poor modified Barthel index at discharge, and extensive cord involvement were associated with severe residual disability. On multivariate analysis, delayed initiation of methylprednisolone was identified as a poor prognostic factor. CONCLUSION: A variety of inflammatory, infective, demyelinating, and autoimmune disorders present with acute transverse myelopathy. Early institution of methylprednisolone reduces the disability in these patients.

Haemophilus influenzae Type a Meningitis in Immunocompetent Child, Oman, 2015.

Meningitis caused by Haemophilus influenzae type b (Hib) was eliminated in Oman after the introduction of Hib vaccine in 2001. However, a case of H. influenzae type a meningitis was diagnosed in a child from Oman in 2015, which highlights the need to monitor the incidence of invasive non-Hib H. influenzae disease.