Nebulized Recombinant Tissue Plasminogen Activator (rt-PA) for Acute COVID-19-Induced Respiratory Failure: An Exploratory Proof-of-Concept Trial.

Acute lung injury in COVID-19 results in diffuse alveolar damage with disruption of the alveolar-capillary barrier, coagulation activation, alveolar fibrin deposition and pulmonary capillary thrombi. Nebulized recombinant tissue plasminogen activator (rt-PA) has the potential to facilitate localized thrombolysis in the alveolar compartment and improve oxygenation. In this proof-of-concept safety study, adults with COVID-19-induced respiratory failure and a <300 mmHg PaO2/FiO2 (P/F) ratio requiring invasive mechanical ventilation (IMV) or non-invasive respiratory support (NIRS) received nebulized rt-PA in two cohorts (C1 and C2), alongside standard of care, between 23 April-30 July 2020 and 21 January-19 February 2021, respectively. Matched historical controls (MHC; n = 18) were used in C1 to explore efficacy. Safety co-primary endpoints were treatment-related bleeds and <1.0-1.5 g/L fibrinogen reduction. A variable dosing strategy with clinical efficacy endpoint and minimal safety concerns was determined in C1 for use in C2; patients were stratified by ventilation type to receive 40-60 mg rt-PA daily for ≤14 days. Nine patients in C1 (IMV, 6/9; NIRS, 3/9) and 26 in C2 (IMV, 12/26; NIRS, 14/26) received nebulized rt-PA for a mean (SD) of 6.7 (4.6) and 9.1(4.6) days, respectively. Four bleeds (one severe, three mild) in three patients were considered treatment related. There were no significant fibrinogen reductions. Greater improvements in mean P/F ratio from baseline to study end were observed in C1 compared with MHC (C1; 154 to 299 vs. MHC; 154 to 212). In C2, there was no difference in the baseline P/F ratio of NIRS and IMV patients. However, a larger improvement in the P/F ratio occurred in NIRS patients (NIRS; 126 to 240 vs. IMV; 120 to 188) and fewer treatment days were required (NIRS; 7.86 vs. IMV; 10.5). Nebulized rt-PA appears to be well-tolerated, with a trend towards improved oxygenation, particularly in the NIRS group. Randomized clinical trials are required to demonstrate the clinical effect significance and magnitude.

Novel formulation with essential oils as a potential agent to minimize African swine fever virus transmission in an in vivo trial in swine.

BACKGROUND AND AIM: African swine fever (ASF) is currently the most prevalent disease in swine. The disease is spreading throughout primary swine-producing countries with heavy losses in population and revenue. To date, no successful vaccines or medications have been reported. This study aimed to design and develop a blend of natural essential oils and test its efficacy against the ASF virus (ASFV) in swine. MATERIALS AND METHODS: We attempted to develop a natural oil blend formulation (NOBF) and determine its efficacy against the ASFV. This study follows on from a previously published in vitro study that reported that the NOBF has anti-ASFV properties. A study was designed using 21 healthy piglets of triple-cross (Landrace + Yorkshire + Durok) crossbred pathogen-free pigs with an average weight of 15 kg. The study consisted of NOBF-incubated, NOBF, positive control, and negative control groups. The NOBF groups were administered NOBF (80 mL/ton mixed in drinking water) beginning 10 days before the challenge and continuing throughout the experiment. The positive and negative control pigs consumed regular drinking water. The pigs were challenged by a sublethal dose of pure isolate ASFV strain Vietnam National University of Agriculture-ASFV-L01/HN/04/19 inoculation with 103.5 HAD50/dose through the intramuscular route. There were sic pigs in each group, three pigs directly IM challenged, and three pigs were considered cohoused pigs. RESULTS: Both challenged (three) and cohoused (three) pigs in the positive control showed clinical signs of ASFV infection, as detected by real-time polymerase chain reaction (RT-PCR) in blood samples, oral swabs, and feces. There was 100% cumulative mortality, that is, both challenged and contact pigs died in the positive control group on day 20 of infection. No signs of infection or mortality were observed in the NOBF-incubated group. The challenged pigs in the NOBF-direct challenge group showed clinical signs and mortality, whereas no clinical signs or symptoms occurred in the cohoused pigs. The immunoglobulin G (IgG) level of the contact pigs was the highest in the treatment group and the lowest in the positive control group. The IgM level of the contact pigs in the treatment groups was the lowest, whereas that of the positive control was the highest. The RT-PCR test showed that the ASFV was deactivated in the NOBF-incubated group. The challenged and contact pigs of the positive control group had high Ct values. The challenged pigs of the NOBF group had high Ct values, whereas the contact pigs from the same group and those of the negative control were negative for the ASFV, determined by PCR, in all samples. The comparison of the challenged groups showed that the appearance of the virus was delayed by at least 2 days in the NOBF group compared to the positive control group. CONCLUSION: The results showed that NOBF can prevent the spread of the ASFV in a population. Moreover, NOBF can enhance the pig humoral immune system by enhancing IgG levels and reducing IgM levels. This study successfully demonstrated that NOBF is an anti-ASFV agent, which prevents horizontal transmission and enhances pig humoral immunity.

Natural oil blend formulation as an anti-African swine fever virus agent in in vitro primary porcine alveolar macrophage culture.

BACKGROUND AND AIM: African swine fever is one of the severe pathogens of swine. It has a significant impact on production and economics. So far, there are no known remedies, such as vaccines or drugs, reported working successfully. In the present study, the natural oil blend formulation's (NOBF) efficacy was evaluated against ASFV in vitro using porcine alveolar macrophages (PAMs) cells of swine. MATERIALS AND METHODS: The capacity of NOBF against the ASFV was tested in vitro. The NOBF combines Eucalyptus globulus, Pinus sylvestris, and Lavandula latifolia. We used a 2-fold serial dilution to test the NOBF formulation dose, that is, 105 HAD50/mL, against purified lethal dose of African swine in primary PAMs cells of swine. The PAM cells survival, real-time polymerase chain reaction (PCR) test, and hemadsorption (HAD) observation were performed to check the NOBF efficacy against ASFV. RESULTS: The in vitro trial results demonstrated that NOBF up to dilution 13 or 0.000625 mL deactivates the lethal dose 105 HAD50 of ASFV. There was no HAD (Rosetta formation) up to dilution 12 or 0.00125 mL of NOBF. The Ct value obtained by running real-time PCR of the NOBF group at 96 h post-infection was the same as the initial value or lower (25), whereas the Ct value of positive controls increased several folds (17.84). CONCLUSION: The in vitro trial demonstrated that NOBF could deactivate the ASFV. The NOBF has the potential to act as anti-ASFV agent in the field. The next step is to conduct in vivo level trial to determine its efficacy.

Treatment of COVID-19: a review of current and prospective pharmacotherapies.

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread and have grave health and socioeconomic consequences worldwide. Researchers have raced to understand the pathophysiological mechanisms underpinning the disease caused by SARS-CoV-2 so that effective therapeutic targets can be discovered. This review summarises the key pharmacotherapies that are being investigated for treatment of COVID-19, including antiviral, immunomodulator and anticoagulation strategies.

Spectrum of gastrointestinal involvement in Stevens - Johnson syndrome.

Stevens - Johnson syndrome (SJS) or toxic epidermal necrolysis (TEN) is a severe adverse drug reaction associated with involvement of skin and mucosal membranes, and carries significant risk of mortality and morbidity. Mucus membrane lesions usually involve the oral cavity, lips, bulbar conjunctiva and the anogenitalia. The oral/anal mucosa and liver are commonly involved in SJS or TEN. However, intestinal involvement is distinctly rare. We herein review the current literature regarding the gastrointestinal involvement in SJS or TEN. This review focuses mainly on the small bowel and colonic involvement in patients with SJS or TEN.

Determining the surface dilational rheology of surfactant and protein films with a droplet waveform generator.

Understanding rheological properties of surfactant and protein films plays a crucial role in a variety of industrial and research areas, such as food processing, cosmetics, and pharmacology. To determine the surface dilational modulus using drop shape analysis, one needs to measure the dynamic surface tension in response to a sinusoidal oscillation of the surface area of the droplet. Despite many applications of drop shape analysis in studying interfacial rheology, oscillation of the droplet surface area is usually controlled in an indirect manner. Existing methods are only capable of controlling volume oscillations of the droplet rather than its surface area. We have developed an arbitrary waveform generator (AWG) to directly oscillate the surface area of a millimeter-sized droplet in a predefined sinusoidal waveform. Here, we demonstrated the capacity of this AWG, in conjunction with constrained drop surfactometry (CDS), in studying the surface dilational rheology of adsorbed surfactant and protein films. It is found that the surface dilational modulus determined for a dilute surfactant (C12DMPO) and two protein solutions (bovine serum albumin and ß-casein) revealed their adsorption mechanisms. Our methods hold promise in studying the interfacial rheology of various thin-film materials, biomembranes, foams, and emulsions.

An expedient approach to pyrrolo[3,2-c]quinolines via regioselective formation of the pyrrole nucleus over indoles.

An efficient approach for the synthesis of pyrrolo[3,2-c]quinolines (the core nucleus of the natural product martinellic acid) from protected 2-alkynylanilines via the regioselective formation of pyrroles followed by Heck and intramolecular Michael addition has been described.

Stereoselective tandem synthesis of thiazolo fused naphthyridines and thienopyridines from o-alkynylaldehydes via Au(III)-catalyzed regioselective 6-endo-dig ring closure.

An operationally simple approach for the stereoselective tandem synthesis of novel thiazolo fused naphthyridines and thienopyridines by the reaction of o-alkynylaldehydes with L-cystine methyl ester hydrochloride via Au(III)-catalyzed regioselective 6-endo-dig ring closure under mild reaction conditions is described. It is noteworthy that alkynes bearing an alkyl and a strong electron-withdrawing nitro group successfully afforded the desired products in good yields.

Palladium-catalyzed regioselective [3 + 2] annulation of internal alkynes and iodo-pyranoquinolines with concomitant ring opening.

A regioselective tandem synthesis of highly functionalized pyrrolo[1,2-a]quinolines has been developed through a novel strategy by palladium-catalyzed [3 + 2] annulation of iodo-pyranoquinolines and internal alkynes with subsequent ring opening. Pyranoquinoline with n-alkyl substitution at the 3-position leads to the formation of pyrrolo-acridones via [3 + 2] annulations/ring opening and successive intramolecular cross-aldol condensation.

Copper-catalyzed tandem synthesis of indolo-, pyrrolo[2,1-a]isoquinolines, naphthyridines and bisindolo/pyrrolo[2,1-a]isoquinolines via hydroamination of ortho-haloarylalkynes followed by C-2 arylation.

An efficient approach for the copper-catalyzed regioselective tandem synthesis of diversely substituted indolo[2,1-a]isoquinolines 11a-r, pyrrolo[2,1-a]isoquinolines 12a-d, and indolo-, pyrrolo[2,1-f][1,6]naphthyridines 14a-f via preferential addition of the heterocyclic amines onto the ortho-haloarylalkynes over N-arylation followed by intramolecular C-2 arylation is described. The scope of the developed chemistry was successfully extended for the direct synthesis of bisindolo-, pyrrolo[2,1-a]isoquinolines 15a-g, a regioisomer of the bisindolo[1,2-a]quinolines used as organic single-crystal field-effect transistor. Hydroxymethyl benzotriazole, which is an inexpensive and air stable compound, has been used as a ligand to carry out this one-step conversion of simple, readily available starting materials into an interesting class of heterocyclic compounds.

Protection of Procambarus clarkii against white spot syndrome virus using recombinant oral vaccine expressed in Pichia pastoris.

The potential for oral vaccination of crayfish against white spot syndrome virus was investigated. The envelope proteins VP19 and VP28 were expressed in yeast (Pichia pastoris). The expressed proteins were used as oral vaccines in different forms viz., in whole culture form, whole culture sonicated form, whole culture centrifuged supernatant form, and cell residue form. The recombinant proteins were mixed with food pellets and fed to crayfish for 25 days. The vaccinated groups were divided into two even groups and challenged on the 3rd and 21st day of post vaccination. Among different vaccine groups the relative percent survival (RPS) values of sonicated form and supernatant form vaccines were found the best and met the criterion (>RPS 60%) of effective vaccine even after 21st day of post vaccination. Development of vaccine by using recombinant proteins VP19 and VP28 in yeast as expression vector was feasible with significant effects.

The efficacy of recombinant vaccines against white spot syndrome virus in Procambarus clarkii.

The effectiveness of oral, mock-, and immersion vaccination was investigated against white spot syndrome virus (WSSV) in crayfish. The most exposed WSSV envelope proteins VP19 and VP28 were used in different compositions and with different modes of applications. In experiment 1 crayfish were fed recombinant protein coated food pellets for 25 days, in experiment 2 the purified proteins were directly injected to them followed by one booster dose on 5th day and in experiment 3 the crayfish were left immersed in vaccines for 7 h. Experimental crayfish were challenged on 3rd and 21st days after last vaccination. The overall result showed that VP28 group has lowest cumulative mortality percentage accounting 39.6% at 3rd day and 39.83% at 21st day when injected and it was 43.2% and 49% when fed orally and 46.3% and 46.5% when immersed at 3rd and 21st days, respectively (p<0.05). In VP19 and VP28 (50:50) mixture, mock vaccination showed better performance (36.5%) over immersion (53%) and oral vaccination (53.2%) when challenged on 3rd day and mock vaccination (36.50%) followed by oral (51%) and immersion vaccination (56.5%) on 21st day as well. The VP19 recombinant mock vaccination group performed better (52.91% of 3rd day and 56.46% of 21st day) than oral (76% of 3rd day and 82% of 21st day) and immersion (83% of 3rd day and 86.3% of 21st day) vaccine groups. All the experimental groups except VP19 were significantly different (p<0.05) from the control groups. A positive cumulative effect was observed when VP28 was mixed with VP19 in equal proportion in all the experimental trials, which shows the effectiveness of VP19 as a vaccine component too. In the present trial on the basis of cumulative mortality percentage it is found that mock-vaccination group is more effective than the oral vaccination and immersion vaccination. It also suggests that specific memory exists in crayfish and the effects of VP28 are significant. The effect of VP19 along with VP28 has also shown significant effect against WSSV.

The LMA Fastrach facilitates fibreoptic intubation in oral cancer patients.

PURPOSE: To compare ease of endotracheal intubation with the Intubating Laryngeal Mask Airway (ILMA) tracheal tube (TT; for LMA-Fastrach) and regular PVC TT (Portex) for nasotracheal fibreoptic intubation in oral cancer patients with a difficult airway. METHODS: 40 patients of physical status ASA I-II with a history of previous oral cancer surgery and/or postoperative radiotherapy scheduled for oral cancer surgery were randomly allocated by sealed envelopes to undergo tracheal intubation with either the ILMA TT or a standard TT. Ease of nasal passage of the TT and ease of tracheal intubation over the fibrescope was assessed. Peak airway pressures were assessed intraoperatively and postoperatively for 12 hr. RESULTS: The use of the ILMA TT increased the ease of nasotracheal intubation by increasing the percentage of successful tube placements at the first attempt (80%) in comparison with standard TT (35%); (P < 0.05). Peak airway pressures were found to remain low with the ILMA TT. None of the patients experienced any airway related complications. CONCLUSIONS: Use of a soft, flexible, nonkinking ILMA TT with a tapered tip design facilitates passage into the trachea over a fibreoptic bronchoscope and allows maintenance of lower airway pressures. The ILMA TT may be a useful adjunct for management of the difficult airway in oral cancer surgery.

Production of recombinant enveloped structural proteins from the Chinese WSSV isolate.

The white spot syndrome virus (WSSV) is one of the deadly pathogens of penaeid shrimps and other crustaceans. The WSSV virion consists of an enveloped rod-shaped nucleocapsid enclosing a large circular double stranded DNA genome of 305 Kb with 181 open reading frames. The two major structural genes, VP19 and VP28 were amplified from the genomic DNA of Chinese isolate of WSSV and cloned in pUCm-T vector and sub cloned in pET-30a (+) vector. The expressions of genes inE. coli (BL21) were confirmed by SDS-PAGE analysis. The clones were sequenced, submitted to the gene bank and the Xiang Shan strain of WSSV were compared with the previous reported sequence of WSSV of various regions which revealed that VP19 and VP28 gene sequences had certain differences from the sequences of similar genes of the isolate already reported. The recombinant proteins expressed, purified and characterized.