Fifty years of sheep red blood cells to monitor humoral immunity in poultry: a scientometric evaluation.

Sheep red blood cells (SRBC) are commonly employed by scientists to address humoral immune responses in poultry. While SRBC are closely related to the study of humoral immunity in poultry, the initial purpose of much research did not focus on the mechanisms involved. Here, we provide a qualitative approach and utilize scientometric techniques, including trend analyses, scientific collaborations and mapping, and word co-occurrence evaluations, to summarize the role of SRBC in the poultry studies. First, a search strategy on Web of Science (WoS) was conducted to find publications that included SRBC in the poultry studies. Publications were partitioned into 4 categories: nutrition, genetics, microbiology, and physiology. For scientometric evaluation, scientific maps and networks were produced to clarify the occurrence of SRBC in the poultry studies. Data used included 702 publications over a period of 50 y (1968-2018) that were retrieved from the WoS database. About 95% of the publications were published in English language. Indigenous, experimental, and commercial chickens, quail, and medicinal plants field/topics were the main subjects of publications. In recent years, authors have used SRBC to study humoral immune response as a secondary aim of their research, especially when poultry production/performance was studied. This was especially the case in recent decades for studies in poultry nutrition. Analysis of keywords co-occurrence showed that the phrase SRBC mostly occurred with chickens, immune response, and especially with broilers. Moreover, the "medicinal plants" are becoming important especially for research on broilers and the reduced use of antibiotics in feed. Consequently, in addition to studying the medicinal plants, finding antibiotic replacements, and/or growth performance in the birds, humoral immunity is suggested to be investigated using SRBC. Moreover, interdisciplinary studies with the cooperation of scientists from agriculture, veterinary, immunology, biochemistry and molecular biology, and toxicology will develop in the future.

Saxifraga spinulosa-Derived Components Rapidly Inactivate Multiple Viruses Including SARS-CoV-2.

Novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus (IAV), and norovirus (NV) are highly contagious pathogens that threaten human health. Here we focused on the antiviral potential of the medicinal herb, Saxifraga spinulosa (SS). Water-soluble extracts of SS were prepared, and their virus-inactivating activity was evaluated against the human virus pathogens SARS-CoV-2 and IAV; we also examined virucidal activity against feline calicivirus and murine norovirus, which are surrogates for human NV. Among our findings, we found that SS-derived gallocatechin gallate compounds were capable of inactivating all viruses tested. Interestingly, a pyrogallol-enriched fraction (Fr 1C) inactivated all viruses more rapidly and effectively than did any of the component compounds used alone. We found that 25 µg/mL of Fr 1C inactivated >99.6% of SARS-CoV-2 within 10 s (reduction of ≥2.33 log10 TCID50/mL). Fr 1C resulted in the disruption of viral genomes and proteins as determined by gel electrophoresis, electron microscopy, and reverse transcription-PCR. Taken together, our results reveal the potential of Fr 1C for development as a novel antiviral disinfectant.

Analysis of the water-soluble polysaccharides from Camellia japonica pollen and their inhibitory effects on galectin-3 function.

Pollen has been defined as dietary supplement used to supplement the diet in many countries, but the primary structure and activity of Camellia japonica pollen polysaccharide remain unclear. In this study, the water-soluble polysaccharide extracted from Camellia japonica pollen (WCPP) was fractionated into one neutral fraction (WCPP-N) and two acidic fractions (WCPP-A1 and WCPP-A2) by DEAE-cellulose column, and WCPP-A2 was further fractionated into two homogeneous sub-fractions (WCPP-A2a and WCPP-A2b) by Sepharose CL-6B column. Monosaccharide composition results showed that WCPP-N might mainly contain starch-like glucan as well as some arabinogalactan, while WCPP-A1, WCPP-A2 and its sub-fractions might mainly composed of rhamnogalacturonan I (RG-I) pectic polysaccharide domain backbone with some different types of side chains, including arabinan, galactan, and/or arabinogalactan. The primary structure analysis of WCPP-A2a by NMR spectra analysis suggested that WCPP-A2a was an RG-I-like pectic polysaccharide, branched at the O-4 of Rha residues in the backbone, with α-(1 â†’ 3,5)-L-arabinan as well as type-II arabinogalactan side chain to which were attached. The results of galectin-3-mediated hemagglutination assay indicated that WCPP-A2a exhibited the strongest inhibitory effect on galectin-3 with MIC value around 0.27 µg/mL. These results suggested the potential use of Camellia japonica pollen polysaccharide as a galectin3 inhibitor in functional foods.

Antiviral activity of Isatidis Radix derived glucosinolate isomers and their breakdown products against influenza A in vitro/ovo and mechanism of action.

ETHNOPHARMACOLOGICAL RELEVANCE: Isatidis Radix, the sun-dried roots of Isatis indigotica Fortune ex Lindl., is one of the most usually used traditional Chinese medicines. For centuries, the herb has been employed in clinical practice for treatment of virus infection and inflammation. However, its active ingredients remain unclear. AIM OF THE STUDY: In the present study, the anti-influenza virus activity of epiprogoitrin, progoitrin, epigoitrin and goitrin, the Isatidis Radix derived glucosinolate isomers and their breakdown products, was firstly evaluated in vitro and in ovo and their mechanism of action was investigated. MATERIALS AND METHODS: Epiprogoitrin, progoitrin, epigoitrin and goitrin were isolated from Isatidis Radix by chiral separation. In vitro and in ovo evaluations were performed on Madin-Darby canine kidney (MDCK) cells and embryonated eggs respectively, both using protocols including prevention, treatment and virus neutralization. Hemagglutination (HA) and neuraminidase (NA) inhibition assays were performed for further understanding of the antiviral mechanism. RESULTS: Isatidis Radix derived glucosinolate isomers and their breakdown products all exhibited dose-dependent inhibition effect against influenza A virus (H1N1) without toxicity. The antiviral potency of the components was in the order of progoitrin > goitrin > epigoitrin > epiprogoitrin. The attachment of the constituents to the viral envelope conduced to the mechanism of their antiviral action without disturbing viral adsorption or budding. CONCLUSION: Taken together, these results are promising for further development of Isatidis Radix and may contribute an adjunct to pharmacotherapy for influenza virus infection.

Antiviral activities of Cholistani plants against common poultry viruses.

Herbal medicines are becoming more popular and acceptable day by day due to their effectiveness, limited side effects, and cost-effectiveness. Cholistani plants are reported as a rich source of antibacterial, antifungal, antiprotozoal, antioxidant, and anticancer agents. The current study has evaluated antiviral potential of selected Cholistani plants. The whole plants were collected, ground and used in extract formation with n-hexane, ethyl acetate and n-butanol. All the extracts were concentrated by using a rotary evaporator and concentrate was finally dissolved in an appropriate vol of the same solvent. All of the extracts were tested for their antiviral potential by using 9-11 days old chick embryonated eggs. Each extract was tested against the Avian Influenza virus H9N2 strain (AIV), New Castle Disease virus Lasoota strain (NDV), Infectious bronchitis virus (IBV) and an Infectious bursal disease virus (IBDV). Hemagglutination test (HA) and Indirect Hemagglutination (IHA) tests were performed for different viruses. The overall order of the antiviral potential of Cholistani plants against viruses was NDV>IBV>IBDV>AIV. In terms of antiviral activity from extracts, the order of activity was n-butanol>ethyl acetate>n-hexane. The medicinal plants Achyranthes aspera, Neuroda procumbens, Panicum antidotale, Ochthochloa compressa and Suaeda fruticose were very effective against all four poultry viruses through their extracts. The low IC50 values of these extracts confirm the high antiviral potential against these viruses. It is worth to mention that Achyranthes aspera was found positive against IBDV through all its extracts which overcome the problem of unavailability of any known drug against IBDV. In short, the study proved that Cholistani plants are rich source of antiviral agent and their extracts can be used as good source of antiviral drugs both in crude and in purified form.

Antiviral Activity of Fritillaria thunbergii Extract against Human Influenza Virus H1N1 (PR8) In Vitro, In Ovo and In Vivo.

Influenza viruses cause respiratory diseases in humans and animals with high morbidity and mortality rates. Conventional anti-influenza drugs are reported to exert side effects and newly emerging viral strains tend to develop resistance to these commonly used agents. Fritillaria thunbergii (FT) is traditionally used as an expectorant for controlling airway inflammatory disorders. Here, we evaluated the therapeutic effects of FT extracts against influenza virus type A (H1N1) infection in vitro, in ovo, and in vivo. In the post-treatment assay, FT extracts showed high CC50 (7,500 µg/ml), indicating low toxicity, and exerted moderate antiviral effects compared to oseltamivir (SI 50.6 vs. 222) in vitro. Antiviral activity tests in ovo revealed strong inhibitory effects of both FT extract and oseltamivir against H1N1 replication in embryonated eggs. Notably, at a treatment concentration of 150 mg/kg, only half the group administered oseltamivir survived whereas the FT group showed 100% survival, clearly demonstrating the low toxicity of FT extracts. Consistent with these findings, FT-administered mice showed a higher survival rate with lower body weight reduction relative to the oseltamivir group upon treatment 24 h after viral infection. Our collective results suggest that FT extracts exert antiviral effects against influenza H1N1 virus without inducing toxicity in vitro, in ovo or in vivo, thereby supporting the potential utility of FT extract as a novel candidate therapeutic drug or supplement against influenza.

Isolates from Monechma ciliatum seeds' extract hampered Porphyromonas gingivalis hemagglutinins.

Porphyromonas gingivalis is a major periodontitis pathogen that produces several virulence factors including hemagglutinins. These proteins, which are vital molecules, allow P. gingivalis to uptake iron and heme by attaching, aggregating, and lysing erythrocytes. In this study, we evaluated the inhibitory activity of the aqueous extract of Monechma ciliatum seeds against the hemagglutination activity of P. gingivalis. M. ciliatum is a Sudanese medicinal herb that grows in arid and semi-arid lands of tropical Africa. The water extracted from dry powdered seeds was partitioned using ethyl acetate followed by reversed-phase chromatography, thin-layer chromatography, ESI-MS, and NMR analysis resulting in the isolation of four compounds identified as oleic acid, coumarin, 1,2-dioleoylglycerol, and 1,3-dioleoylglycerol with MICs of 15-100 µg/ml against hemagglutination. We believe that the isolation and purification of these compounds will expand the application of M. ciliatum as a natural therapeutic or preventative agent. PRACTICAL APPLICATIONS: Monechma ciliatum or black mahlab is a famous medicinal plant that grows in some parts of arid and semi-arid areas of tropical Africa including western Sudan. Despite its nutritional and traditional medical applications, no studies have evaluated its anti-hemagglutination activity against periodontal pathogens. In this study, four active compounds (oleic acid, coumarin, 1,2-dioleoylglycerol, and 1,3-dioleoylglycerol) were isolated and identified from an aqueous extract of M. ciliatum seeds. The isolated compounds revealed high levels of inhibitory activity against all hemagglutinin agents secreted by Porphyromonas gingivalis. This evidence of inhibitory activity will encourage the application of M. ciliatum effectively as a functional food or therapeutic agent to prevent periodontal diseases in the early stages.

Green synthesis, characterisation and biological evaluation of plant-based silver nanoparticles using Quercus semecarpifolia Smith aqueous leaf extract.

The development of reliable and green methods for the fabrication of metallic nanoparticles (NPs) has many advantages in the field of nanotechnology. In this direction, the present work describes an eco-friendly and cost-effective protocol for the production of silver NPs (AgNPs) using an aqueous extract of Quercus semecarpifolia leaves. Different techniques were carried out for the characterisation of the synthesised AgNPs. The ultraviolet-visible spectroscopic analysis showed the highest absorbance peak at 430 nm. The particle size and structure were confirmed by scanning electron microscopy as well as transmission electron microscopy (TEM) analysis. From TEM imaging, it was revealed that the formed particles were spherical with an average size of 20-50 nm. The crystalline nature of the NPs was determined by X-ray powder diffraction patterns. Thermogravimetry and differential thermal analysis were also evaluated by a temperature increment from 100 to 1000°C. Bio-inspired synthesis of AgNPs was performed for their pharmacological evaluation in relation to the activities of the crude methanolic, n-hexane, chloroform, ethyl acetate, and aqueous extracts. Good cytotoxic activity was exhibited by the green-synthesised AgNPs (77%). Furthermore, the AgNPs were found to exhibit significant antioxidant activity at 300 µg/ml (82%). The AgNPs also exhibited good phytotoxic potential (75%).

REPORT- Some ethanobotanically important plants from Cholistan area for anti avian influenza virus (AIV) H9N2 screening.

Avian influenza or bird flu is a common problem of domestic and wild birds. Some of its strains are able to cross the species barrier and cause infection in various members of class Mammalia. In view of relatively lesser efficacy of vaccines, antiviral therapies remain the only choice for the sustenance of mammals acquiring this highly devastating infection. This study is based on the evaluation of antiviral potential of methanol extracts of eleven selected Cholistani plants. The methanol extracts were prepared by using dried plants material followed by concentrating in a rotary evaporator and finally air dried before dissolving in nanopure water. The suspension was filter sterilized and subjected to in ovo antiviral assays. The allantoic fluids were harvested and haemagglutinin (HA) titers were determined. Among the eleven plants evaluated all methanol extracts were found effective against AIV H9N2 except S. baryosma extract. The medicinal plants O. compressa, N. procumbens, and S. surattense were found to be more effective than others and they retained HA titers at 0 after challenge. The next in order were extracts of O. esculentum, H. salicornicum and S. fruticosa which kept HA titers at 4, 8 and 16 respectively. The extracts of H. recurvum, P. antidotale, S. icolados and A. aspera were found less effective than above mentioned plant extracts and they kept the HA titers at 32, 64, 128 and 256 respectively. These results led us to conclude that the medicinal plants of Cholistan region are a rich source of antiviral agent(s) against AIV H9N2 and could be a source of cost effective alternate therapeutics.

Isolation and Characterization of Lectin from the Artist's Conk Medicinal Mushroom, Ganoderma applanatum (Agaricomycetes), and Evaluation of Its Antiproliferative Activity in HT-29 Colon Cancer Cells.

The growth and lectin production of Ganoderma applanatum, a white rot fungus, was optimized in broth cultures. The fungus was found to have a higher growth rate and higher lectin activity when grown in a medium adjusted to pH 6.5 at 26°C under stationary conditions. Expression of lectin activity started in 5-day-old mycelial culture; maximum activity was expressed after the 15th day of incubation. Among the various carbon and nitrogen sources tested, the carbon source sucrose and the nitrogen source yeast extract support maximum growth and lectin production. Lectin from G. applanatum was purified by ammonium sulfate precipitation and ion exchange chromatography. The purified fraction revealed a single band with a molecular weight of 35.0 kDa. Moreover, carbohydrates such as mannitol, glucose, sucrose, maltose, mannose, galactose, sorbose, and fructose were found to inhibit the hemagglutinating activity of the lectin. The purified lectins from G. applanatum contain cytotoxic and proapoptotic activities against HT-29 colon adenocarcinoma cells.

Antibacterial, antioxidant and Immuno-modulatory properties in extracts of Barleria lupulina Lindl.

BACKGROUND: Antibacterial, immunomodulatory and antioxidant properties of aerial parts of Barleria lupulina Lindl were investigated in the present communication. METHODS: The antibacterial, antioxidant and immunomodulatory properties of B. lupulina (methanol soluble leaf and stem extracts) was analyzed by minimum inhibitory concentration, total phenolic contents, DPPH radical scavenging activity, determination of toxicity, hemagglutination antibody titre, delayed type hypersensitivity and neutrophil adhesion test, respectively. RESULTS: Methanol soluble leaf extract (MLE) contains more soluble bioactive compounds inhibiting the growth of five bacterial pathogens viz., Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Salmonella typhi even at MICs of 1.25 and 2.5 mg/mL. Aqueous stem extract (ASE) was least effective while MLE was highly effective in inhibiting the growth and survival of bacterial pathogens. While testing the effect of these extracts in animal model, no mortality of albino rats was recorded by using MLE and MSE at the concentrations from 200 to 600 mg/kg of their body weight. The MLE showed significant result in agglutination reaction and induced paw edema volumes when compared with untreated group (control). Both MLE and MSE extracts significantly increased neutrophil adhesion with increase in doses of extracts. MLE was found to have more potent immune-stimulant properties than the MSE. High phenolic contents were found in MSE while lowest IC50 values were found in MLE in term of DPPH radical scavenging activity. CONCLUSIONS: Methanol soluble leaf and stem extract of Barleria lupulina contains antibacterial, antioxidants and immunomodulating phytochemical compounds that was effective for antibacterial, antioxidant and immunomodulatory properties. These may be used as synthetic drug.

Antioxidant and Biological Activities of Proteinaceous Extract from Algerian Glycine max Plant.

OBJECTIVE: Glycine max is commonly used in Algeria for treatment of anemia deficiency and osteoporosis, it ranks first in terms of vegetal proteins. The experiment was aimed at characterizing the proteinaceous Glycine max extract and evaluating its antioxidant, biological and hematological potential. METHODOLOGY: Extraction of proteinaceous materials from Glycine max plant was undertaken using water and n-hexane as extracting media. The isolation of proteins from the crude materials was done, providing the use of ammonium sulfate. The Glycine max proteins were characterized by UV-visible and FT-IR spectroscopy and analyzed by SEM micrograph and x-ray diffraction (XRD). Rheological parameters G' and G'' were assessed. The isolated proteins were tested for their antioxidant, antimicrobial and hemagglutination activities. RESULTS: There was a gelling effect of the protein extract which can be used as an alternative in principally made vaccines with its microbiological and antifungal activities. CONCLUSION: The proteinaceous extract from Algerian Glycine max would have a potential use in biomedical application.

Response induced in Mycoplasma gallisepticum under heat shock might be relevant to infection process.

Despite the fact the term "proteome" was proposed to characterize a set of proteins in one of mycoplasma species, proteome response to various exposures in this bacteria are still obscure. Commonly, authors studying proteomic response on perturbation models in mycoplasmas use single approach and do not confirm their findings by alternative methods. Consequently, the results of proteomic analysis should be validated by complementary techniques. In this study we utilized three complementary approaches (SWATH, MRM, 2D-DIGE) to assess response of Mycoplasma gallisepticum under heat stress on proteomic level and combined these findings with metabolic response and the results of transcriptional profiling. We divide response into two modes - one is directly related to heat stress and other is triggered during heat stress, but not directly relevant to it. The latter includes accumulation of ATP and shedding of antigens. Both of these phenomena may be relevant to evasion of host's immune system and dissemination during mycoplasmosis in vivo.

In vivo antiviral potential of Glycyrrhiza glabra extract against Newcastle disease virus.

Newcastle disease is highly infectious viral disease causing huge economic losses worldwide. These losses can be prevented by control of viral diseases. Medicinal plants have been traditionally used for treatment of different diseases since long. In this study the effect of extracts from Glycyrrhiza glabra leaves are investigated against Newcastle disease virus (NDV) by an in-vivo assay. Seven groups of nine-day-old embryonated chicken eggs were inoculated with various treatments of different plant extracts. All the groups except uninoculated negative control group were inoculated with velogenic NDV strain; five groups received different concentrations of the three extracts. Daily observe the rate of embryo survival. Allantoic fluid from treated eggs was collected for hem agglutination test. Results showed that embryo survival rate was higher 300µg/mL treated group as all the extracts showed antiviral activity. Similarly, the plant extracts effectively control virus as no viruses were identified in the allantoic fluids of all groups treated with low doses of plant. The current results have clearly verified that all the extracts especially that of methanol 300µg/mL from leaves of Glycyrrhiza glabra have strong antiviral activity against NDV in vivo.

Ethanol extract and chromatographic fractions of Tamarindus indica stem bark inhibits Newcastle disease virus replication.

CONTEXT: The plethora of ethnomedicinal applications of Tamarindus indica Linn. (Leguminosae), tamarind, includes treatment of human and livestock ailments; preparations are recognized antipyretics in fevers, laxatives and carminatives. African folklore has various applications of tamarind. However, in Nyasaland, domestic fowl are fed with preparations for prophylactic properties. OBJECTIVES: The objective of this study is to evaluate the antiviral properties of T. indica extract. MATERIALS AND METHODS: Tamarindus indica stem bark was extracted through ethanol maceration over 24 h, and the crude extract was fractionated by gravity-propelled column chromatography. Newcastle disease virus (NDV) inhibitory activity of extract and fractions were evaluated in vivo using 10-d-old embryonated chicken egg (ECE) as the medium for virus cultivation and antivirus assay. About 240 ECE were grouped into eight (three controls and five experimental) and, 200 µL of the extract and fractions respectively inoculated into NDV pre-infected eggs and incubated at 37 °C. Allantoic fluid was harvested 5 d post-virus infection and assayed for haemagglutination (HA). RESULTS: Anti-NDV assessment showed 62.5 mg/mL of crude extract and fractions: TiA, TiC and TiD to yield a HA titre of 1:128 each, while TiB showed 1:64 HA titre. At 125 mg/mL, a titre of 1:16 was recorded against TiB and TiD and, 1:8 against TiA. Similarly, crude extract and TiC, each recorded 1:4 HA titre. However, the minimum concentrations of extract and fraction for virus inactivation were 0.24 mg/mL and 0.49 mg/mL, respectively. CONCLUSION: The antiviral activity shown by T. indica portends novel antiviral drugs and, perhaps, as scaffold for new drugs.

Characterization and functional analysis of a novel C-type lectin from the swimming crab Portunus trituberculatus.

C-type lectins (CTLs) are a family of calcium-dependent carbohydrate-binding proteins. In the present study, a novel C-type lectin (designated as PtCTL1) was identified and characterized from Portunus trituberculatus. The full-length cDNA of PtCTL1 was of 702 bp, containing a 5' untranslated region (UTR) of 91 bp, a 3' UTR of 110 bp with a poly (A) tail, and an open reading frame (ORF) of 501 bp encoding a polypeptide of 166 amino acids with a putative signaling peptide of 21 amino acids. A C-type lectin carbohydrate-recognition domain (CRD) containing four conserved cysteines was identified in the amino acid sequence of PtCTL1. The cDNA fragment encoding the mature peptide of PtCTL1 was recombined into pET-21a(+) with a C-terminal hexa-histidine tag fused in-frame and expressed in Escherichia coli Origami (DE3). The recombinant PtCTL1 (rPtCTL1) can agglutinate all the tested bacteria, including three Gram-positive bacterial strains and three Gram-negative bacterial strains. In addition, erythrocyte agglutination and LPS-binding activity were observed in a Ca2+-dependent manner. The erythrocyte agglutination was inhibited by EDTA, indicating that PtCTL1 was Ca2+-dependent. The mRNA transcripts of PtCTL1 were detected mainly in the tissues of hepatopancreas and hemocytes and its levels were significantly up-regulated in hemocytes following Vibrio alginolyticus challenge. These results indicate that PtCTL1 may function as a pattern recognition receptor (PRR) for protecting P. trituberculatus from bacterial infection. Moreover, such findings also provide evidence for further understanding the innate immunology of invertebrate.

Size distribution analysis of influenza virus particles using size exclusion chromatography.

Size exclusion chromatography is a standard method in quality control of biopharmaceutical proteins. In contrast, vaccine analysis is often based on activity assays. The hemagglutination assay is a widely accepted influenza quantification method, providing no insight in the size distribution of virus particles. Capabilities of size exclusion chromatography to complement the hemagglutination assay are investigated. The presented method is comparatively robust regarding different buffer systems, ionic strength and additive concentrations. Addition of 200mM arginine or sodium chloride is necessary to obtain complete virus particle recovery. 0.5 and 1.0M arginine increase the hydrodynamic radius of the whole virus particles by 5nm. Sodium citrate induces virus particle aggregation. Results are confirmed by dynamic light scattering. Retention of a H1N1v strain correlates with DNA contents between 5ng/mL and 670ng/mL. Quantitative elution of the virus preparations is verified on basis of hemagglutination activity. Elution of hemagglutination inducing compounds starts at a flow channel diameter of 7000nm. The universal applicability is demonstrated with three different influenza virus samples, including an industrially produced, pandemic vaccine strain. Size distribution of the pandemic H1N1v 5258, H1N1 PR/8/34, and H3N2 Aichi/2/68 preparations spreads across inter- and intra-particle volume and extends to the secondary interaction dominated range. Thus, virus particle debris seems to induce hemagglutination. Fragments generated by 0.5% Triton™ X-100 treatment increase overall hemagglutination activity.

Effect of oral cranberry extract (standardized proanthocyanidin-A) in patients with recurrent UTI by pathogenic E. coli: a randomized placebo-controlled clinical research study.

PURPOSE: To evaluate the effect of cranberry extract (PAC-A ~ proanthocyanidin-A) on the in vitro bacterial properties of uropathogenic (E. coli) and its efficacy/tolerability in patients with subclinical or uncomplicated recurrent UTI (r-UTI). MATERIALS AND METHODS: After obtaining clearance from the ethics committee and administering a written informed consent, 72 patients with r-UTI were enrolled as per protocol (November 2011 to March 2013) in this prospective study, to randomly receive (PAC-A: group I, 36) or (placebo: group II, 36), for 12 weeks. Any change/reduction in the incidence of r-UTI at 12 weeks was construed to be the primary endpoint of this study. RESULTS: After 12 weeks, bacterial adhesion scoring decreased (0.28)/(2.14) in group I/II (p < 0.001); 32/36 (88.8 %) and 2/36 (5.5 %) in groups I and II, respectively, turned MRHA negative (p < 0.001); biofilm (p < 0.01) and bacterial growth (p < 0.001) decreased in group I; microscopic pyuria score was 0.36/2.0 in group I/II (p < 0.001); r-UTI decreased to 33.33 versus 88.89 % in group I/II (p < 0.001); mean subjective dysuria score was 0.19 versus 1.47 in group I/II (p < 0.001), while mean urine pH was 5.88 versus 6.30 in group I/II (p < 0.001). No in vitro antibacterial activity of cranberry could be demonstrated, and no adverse events were noted. CONCLUSIONS: The overall efficacy and tolerability of standardized cranberry extract containing (PAC-A) as a food supplement were superior to placebo in terms of reduced bacterial adhesion; bacterial MRHA negativity; urine pH reduction; and in preventing r-UTI (dysuria, bacteriuria and pyuria). Larger randomized controlled trials are needed to elucidate the precise role, exact dose and optimal duration of PAC-A therapy in patients at risk of r-UTI.

Antibacterial, antifungal, phytotoxic, antioxidant and hemagglutination activities of organic fractions of Arisaema tortuosum.

In the current study, the antimicrobial, phytotoxic, haemagglutination and antioxidant potential of crude methanolic extract (Crd. MeOH Ext.) and four organic fractions of Arisaema tortuosum was investigated. All fractions have been screened for antimicrobial properties against eight bacterial pathogens and six fungal pathogens using agar well diffusion and tube dilution method, respectively. Furthermore, the organic fractions were also screened for its phytotoxicity against Lemna minor. Haemagglutination was performed against all human blood groups while free radical scavenging activity was performed to investigate the antioxidant potential of A. tortuosum. Results obtained for antibacterial activity exhibited various degree of zone of inhibition and significant activity was observed for Pseudomonas aeruginosa (27.16±0.60) followed by Bacillus cereus (18.55±0.69) for Crd. MeOH Ext. and chloroform (CHCl3) fraction, respectively while some strains showed resistant at same concentration. Similarly, non-significant antifungal activity was observed for the plant extracts. However, the highest activity among the strains was observed for Alternaria alternata (22±1.24%) and Aspergillus niger (20±1.00%) for ethyl acetate (EtOAc) fraction and Crd. MeOH Ext., respectively. The plant extracts showed good phytotoxic activity with 77.06% inhibition for n-hexane fraction at 1000µg/mL. The result of Nitric Oxide (NO) reducing assay revealed that the plant has less antioxidant activity with 46.06% inhibition for CHCl(3) fraction at 900µg/mL. For haemagglutination assay, the result displayed no agglutination in all the testing concentration. Based on the current results, it can be concluded that A. tortuosum has significant antimicrobial and moderate phytotoxic potential and therefore can leads to antibiotics and herbicide production.

Immunostimulant activities of the aqueous methanolic extract of Leptadenia pyrotechnica, a plant from Cholistan desert.

ETHNOPHARMACOLOGICAL RELEVANCE: Leptadenia pyrotechnica (Forssk.) Decne (Asclepiadaceae) is a well-renowned medicinal shrub, used by herbal practitioners for various ailments including allergic rhinitis, productive cough, abortion, diabetes, stomach disorders, fever, kidney disorders, stones and cancer AIM OF THE STUDY: On the basis of folkloric uses, L. pyrotechnica was selected from the wide medicinal flora of the Cholistan desert of Pakistan, for the exploration of immunomodulatory potential. MATERIALS AND METHODS: Aqueous methanolic (30:70) extract of aerial parts of L. pyrotechnica (Lp. Cr) was prepared by 3 days thrice maceration and subsequent evaporation under reduced pressure. In-vivo experiments were performed in Wistar albino rats including neutrophil adhesion test, haemagglutinating antibody (HA) titre, delayed-type hypersensitivity response, phagocytic activity and cyclophosphamide-induced myelosuppression. RESULTS: Lp. Cr produced a significant increase in phagocytic index in dose-dependent fashion (3.56, 4.18 and 5.42 at 30, 100 and 300mg/kg, respectively) as well as an augmented response in the delayed-type hypersensitivity reaction and HA titre induced by sheep erythrocytes. Lp. Cr also showed improved adhesion of neutrophils with nylon pellets (15.28, 27.85 and 38.42% at the doses of 30, 100 and 300mg/kg) and prevented myelosuppression in cyclophosphamide-treated rats as evidenced from the hematological parameters. The results of treatment were compared with normal and standard control groups throughout the study and the effects by Lp. Cr (300mg/kg) were found to be comparable with Levamisole. CONCLUSIONS: The results indicated that L. pyrotechnica possesses immunostimulatory activity and justify its traditional use for the control and management of diseases in which the immune system needs to be stimulated like infectious diseases.