Honey and Nigella sativa against COVID-19 in Pakistan (HNS-COVID-PK): A multicenter placebo-controlled randomized clinical trial.

Until now, no specific and effective treatment exists for coronavirus disease 2019 (COVID-19). Since honey and Nigella sativa (HNS) have established antiviral, antibacterial, antiinflammatory, antioxidant, and immunomodulatory properties, we tested their efficacy for this disease in a multicenter, placebo-controlled, and randomized clinical trial at four medical care facilities in Pakistan. RT-PCR confirmed COVID-19 adults showing moderate or severe disease were enrolled in the trial. Patients were randomly assigned in a 1:1 ratio to receive either honey (1 g kg-1 day-1 ) and Nigella sativa seeds (80 mg kg-1 day-1 ) or a placebo for up to 13 days along with standard care. The outcomes included symptoms' alleviation, viral clearance, and 30-day mortality in the intention-to-treat population. Three hundred and thirteen patients, 210 with moderate and 103 with severe disease, underwent randomization from April 30 to July 29, 2020. Among the moderate cases, 107 were assigned to HNS, whereas 103 were assigned to the placebo group. Among the severe cases, 50 were given HNS, and 53 were given the placebo. HNS resulted in ~50% reduction in time taken to alleviate symptoms as compared to placebo (moderate cases: 4 vs. 7 days, Hazard Ratio [HR]: 6.11; 95% Confidence Interval [CI]: 4.23-8.84, p < 0.0001 and for severe cases: 6 vs. 13 days, HR: 4.04; 95% CI: 2.46-6.64; p < 0.0001). HNS also cleared the virus earlier than placebo in both moderate cases (6 vs. 10 days, HR: 5.53; 95% CI: 3.76-8.14, p < 0.0001) and severe cases (8.5 vs. 12 days, HR: 4.32; 95% CI: 2.62-7.13, p < 0.0001). HNS further led to a better clinical score on day 6 with normal activity resumption in 63.6% vs. 10.9% among moderate cases (OR: 0.07; 95% CI: 0.03-0.13, p < 0.0001) and hospital discharge in 50% versus 2.8% in severe cases (OR: 0.03; 95% CI: 0.01-0.09, p < 0.0001). In severe cases, the mortality rate was less than 1/4th in the HNS group than in placebo (4% vs. 18.87%, OR: 0.18; 95% CI: 0.02-0.92, p = 0.029). No HNS-related adverse effects were observed. HNS, compared with placebo, significantly improved symptoms, expedited viral load clearance, and reduced mortality in COVID-19 patients. This trial was registered on April 15, 2020 with ClinicalTrials.gov Identifier: NCT04347382.

Pollen morphology of selected melliferous plants and its taxonomic implications using microscopy.

This research aimed to explore the melliferous bee flora pollen from arid zones of District Attock, Islamabad Capital and Rawalpindi. Morpho-palynological features has provided key information for the classification and help to explain the taxonomy of several genera and species. Therefore, in this work, we examined the morphological characters of pollen potentially to be used for the species delimitation of bee flora. A total of 18 bee floral species, classified into 12 different families were collected, pressed, identified and then acetolyized to observe the grains under microscopy. Bee pollen can be described by small, medium to large sized morphotypes presenting five types of shapes, prolate spheroidal in (seven species), sub-prolate (six species), oblate spheroidal (three species), oblate and prolate (one species each) was examined. The observed polar and equatorial diameter were found maximum in Zea mays 73.5 µm and Pelargonium inquinans 66.75 µm respectively. Pollen of six different types was found namely: tricolporate, 3-zonocolporate, monoportae, tetracolporate, tetraporate and mononsulcate. Whereas highest value of colpi length was measured in Grewia tenax (24.55 µm). Exine surface nature of pollen was examined echinate psilate and scabrate. Exine thickness noted to be maximum in Verbena tenuisecta (8.40 µm) and minimum in Citrus macrocarpa (0.4 µm). Bee floral species considered difficult to identify based on other morphological traits were successfully distinguished using pollen quantitative and qualitative traits, confirming the importance of pollen morphology to diagnose characters to correctly identify honeybee flora. RESEARCH HIGHLIGHTS: Pollen diversity among honeybee flora. Highly variations observed via microscopy in pollen micromorphology. Palyno-morph has taxonomic value for the identification of honeybee floral species.

Prophylactic potential of honey and Nigella sativa L. against hospital and community-based SARS-CoV-2 spread: a structured summary of a study protocol for a randomised controlled trial.

OBJECTIVES: Considering the therapeutic potential of honey and Nigella sativa (HNS) in coronavirus disease 2019 (COVID-19) patients, the objective of the study is defined to evaluate the prophylactic role of HNS. TRIAL DESIGN: The study is a randomized, placebo-controlled, adaptive clinical trial with parallel group design, superiority framework with an allocation ratio of 1:1 among experimental (HNS) and placebo group. An interim analysis will be done when half of the patients have been recruited to evaluate the need to adapt sample size, efficacy, and futility of the trial. PARTICIPANTS: All asymptomatic patients with hospital or community based COVID-19 exposure will be screened if they have had 4 days exposure to a confirmed case. Non-pregnant adults with significant exposure level will be enrolled in the study High-risk exposure (<6 feet distance for >10min without face protection) Moderate exposure (<6 feet distance for >10min with face protection) Subjects with acute or chronic infection, COVID-19 vaccinated, and allergy to HNS will be excluded from the study. Recruitment will be done at Shaikh Zayed Post-Graduate Medical Institute, Ali Clinic and Doctors Lounge in Lahore (Pakistan). INTERVENTION AND COMPARATOR: In this clinical study, patients will receive either raw natural honey (0.5 g) and encapsulated organic Nigella sativa seeds (40 mg) per kg body weight per day or empty capsule with and 30 ml of 5% dextrose water as a placebo for 14 days. Both the natural products will be certified for standardization by Government College University (Botany department). Furthermore, each patient will be given standard care therapy according to version 3.0 of the COVID-19 clinical management guidelines by the Ministry of National Health Services of Pakistan. MAIN OUTCOMES: Primary outcome will be Incidence of COVID-19 cases within 14 days of randomisation. Secondary endpoints include incidence of COVID-19-related symptoms, hospitalizations, and deaths along with the severity of COVID-19-related symptoms till 14th day of randomization. RANDOMISATION: Participants will be randomized into experimental and control groups (1:1 allocation ratio) via the lottery method. There will be stratification based on high risk and moderate risk exposure. BLINDING (MASKING): Quadruple blinding will be ensured for the participants, care providers and outcome accessors. Data analysts will also be blinded to avoid conflict of interest. Site principal investigator will be responsible for ensuring masking. NUMBERS TO BE RANDOMISED (SAMPLE SIZE): 1000 participants will be enrolled in the study with 1:1 allocation. TRIAL STATUS: The final protocol version 1.4 was approved by institutional review board of Shaikh Zayed Post-Graduate Medical Complex on February 15, 2021. The trial recruitment was started on March 05, 2021, with a trial completion date of February 15, 2022. TRIAL REGISTRATION: Clinical trial was registered on February 23, 2021, www.clinicaltrials.gov with registration ID NCT04767087 . FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). With the intention of expediting dissemination of this trial, the conventional formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol. The study protocol has been reported in accordance with the Standard Protocol Items: Recommendations for Clinical Interventional Trials (SPIRIT) guidelines.

Anti-COVID property of subcutaneous ivermectin in synergy with zinc among midlife moderately symptomatic patients: a structured summary of a study protocol for a randomised controlled trial.

OBJECTIVES: The study objective is to quantify the effectiveness of ivermectin (subcutaneous/oral IVM) in the presence or absence of zinc (Zn) for clinical and radiological improvement in coronavirus disease 2019 (COVID-19) patients with moderate severity. TRIAL DESIGN: This quadruple-blinded, placebo-controlled randomized clinical trial will be a multiarmed multi-centered study with superiority framework. PARTICIPANTS: Quinquagenarian and sexagenarian patients with moderate COVID-19 symptoms and positive severe respiratory syndrome coronavirus -2 (SARS-CoV-2) PCR will be included. Participants with co-morbidities and pregnant women will be excluded. Patient recruitment will be done in Shaikh Zayed Medical Complex, Doctors Lounge and Ali Clinic in Lahore (Pakistan). INTERVENTION AND COMPARATOR: The registered patients will be allocated in 6 groups (30 participants each). Patients will be taking subcutaneous IVM at 200 µg/kg/48 h (Arm A) or subcutaneous IVM at 200 µg/kg/48 h and oral Zn 20mg/8 h (Arm B) or oral IVM at 0.2 mg/kg/day (Arm C) or oral IVM at 0.2 mg/kg/day and oral Zn 20mg/8 h (Arm D) or alone oral Zn 20mg/8 h (Arm E) or placebo alone (Arm X). Patients in all arms will receive standard care and respective placebo (empty capsule 8 hourly and/or subcutaneous normal saline 2ml/48 h). MAIN OUTCOMES: Primary endpoints will be duration of symptomatic phase and SARS-CoV-2 clearance along with high resolution CT (HRCT) chest score and clinical grade scale (CGS) on day 6. 30-day mortality will be documented as a secondary endpoint. SARS-CoV-2 clearance will be calculated by second PCR on day 7. HRCT chest score will be measured by the percentage and lung lobes involvement on day 6 with a maximum score of 25. CGS will be recorded on a seven-point scale; grade 1 (not hospitalized, no evidence of infection and resumption of normal activities), grade 2 (not hospitalized, but unable to resume normal activities), grade 3 (hospitalized, not requiring supplemental oxygen), grade 4 (hospitalized, requiring supplemental oxygen), grade 5 (hospitalized, requiring nasal high-flow oxygen therapy and/or noninvasive mechanical ventilation), grade 6 (hospitalized, requiring ECMO and/or invasive mechanical ventilation) and grade 7 (death). RANDOMISATION: A simple lottery method will be used to randomly allocate scrutinized patients in 1:1:1:1:1:1 ratio in 6 groups. BLINDING (MASKING): Patients, primary care physicians, outcome assessors and the data collection team will be blinded. NUMBERS TO BE RANDOMISED (SAMPLE SIZE): 180 participants will be randomized into six arms with five investigational and one placebo group. TRIAL STATUS: Institutional Review Board Shaikh Zayed Post-Graduate Medical Complex, Lahore, Pakistan has approved the protocol (version 2.3) with ID SZMC/IRB/Internal0056/2020. The trial was approved on July 14, 2020, and enrolment started on July 30, 2020. The estimated completion date is October 30, 2021. TRIAL REGISTRATION: Clinical Trial has been retrospectively registered on www.clinicaltrials.gov with registration ID NCT04472585 dated July 16, 2020. FULL PROTOCOL: The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). With the intention of expediting dissemination of this trial, the conventional formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol. The study protocol has been reported in accordance with the Standard Protocol Items: Recommendations for Clinical Interventional Trials (SPIRIT) guidelines.

Microscopic implication and evaluation of herbaceous melliferous plants of southern Khyber Pakhtunkhwa-Pakistan using light and scanning electron microscope.

The aim of the present study is to investigate the pollen morphology of melliferous plant taxa of Southern Khyber Pakhtunkhwa-Pakistan. Using light microscope (LM) and scanning electron microscope (SEM), the palynological study of 18 species of melliferous plants namely Calendula arvensis, Cenchrus pennisetiformis, Citrullus colocynthis, Cucumis melo subsp. agrestis var. agrestis, Cucurbita maxima, Cymbopogon jwarancusa, Cynodon dectylon, Dactyloctenium aegyptium, Helianthus annus, Lagenaria siceraria, Launaea procumbens, Luffa cylindrica, Pennisetum glaucum, Saccharum spontaneum, Sonchus asper, Verbesina encelioides, Xanthium strumarium, and Zea mays was carried out. Both qualitative and quantitative characteristics of pollen were studied. Variations were observed in pollen morphology. The dominant pollen shape was prolate-spheroidal (11 species). All the pollen units were monad. The highest exine thickness was found in Citrullus colocynthis (8.45 µm). The maximum polar and equatorial diameter (102 and 97.55 µm) was found in Luffa cylindrica. Similarly, the highest P/E ratio was found in Cucurbita maxima (1.46). Most of the species showed tricolpate and monoporate type of pollen. The exine sculpturing, number of spines per pollen and between colpi and the pollen fertility and sterility provided significant results for the documentation of melliferous plants. Thus, the information listed in this article will prove helpful to identify the potential melliferous plants in the area, geographical origin of the honey, and the availability of pure honey in the local and international market.

Microscopic investigation of pollen morphology of Brassicaceae from Central Punjab-Pakistan.

The intention of the current study is to provide an account on the palynological features of Brassicaceae from Central Punjab-Pakistan as a basis for future studies. Different morpho-palynological characteristics both qualitative and quantitative were analyzed during this research which includes shape of pollen, diameter of pollen, P/E ratio, exine sculpturing, thickness of exine, type of pollen, shape and size of lumens, and thickness of murus. Taxonomic keys were also constructed based on pollen morphological characters for correct identification of species. This study aims to provide detailed information of pollen diversity and their exine structure based on both qualitative and quantitative characters by using Light microscopy and Scanning electron microscopy. Shape of pollen is mostly prolate, but some species also have sub-prolate to spheroidal prolate types. Exine ornamentation in most species was reticulate, whereas micro reticulate (one species) and coarsely reticulate (one species) exine also observed in some pollen. All the pollen mentioned in this study have tricolpate apertures. Variation found in thickness of exine and other characters proved to be helpful at generic and specific level. The results reinforced the significance of pollen morphological features of family Brassicaceae and aid for valuable taxonomic tool in plant systematics.