Use of the probiotic <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> 299 to reduce pathogenic bacteria in the oropharynx of intubated patients: a randomised controlled open pilot study

Introduction  

Ventilator-associated pneumonia (VAP) is usually caused by aspiration of pathogenic bacteria from the oropharynx. Oral decontamination with antiseptics, such as chlorhexidine (CHX) or antibiotics, has been used as prophylaxis against this complication. We hypothesised that the probiotic bacteria Lactobacillus plantarum 299 (Lp299) would be as efficient as CHX in reducing the pathogenic bacterial load in the oropharynx of tracheally intubated, mechanically ventilated, critically ill patients.     

Not <Emphasis Type="Italic">if</Emphasis> but <Emphasis Type="Italic">when</Emphasis>; no longer <Emphasis Type="Italic">why</Emphasis> but <Emphasis Type="Italic">how</Emphasis>

There is accruing evidence that information technology can improve patient health care, with several trials of technology showing smaller numbers of medication errors, or can provide earlier detection of adverse events. Critics of this type of research point out that better resolution of events is of no value unless their direct management influences clinical outcome. Nevertheless, indirect evidence is available, such as reports indicating the importance of providing specialist neuro-critical care in the management of patients with traumatic brain injury. These studies do not indicate which aspects of critical care management are crucial, but management aimed at the earlier detection and treatment of adverse events must be partly responsible. We continue to hope for definitive controlled trial evidence that information technology-led management yields improved patient outcome, but our experience so far of funding and conducting such studies has been poor. There is no question that we need better monitoring and event detection technology for health care and that we need more research into optimising that technology, but should their adoption depend on large-scale clinical trials? Perhaps now the questions we need to focus upon are no longer if but when, and no longer why but how.     

Development and Evaluation of Probioticated Cucumber Juice Using <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis>

The aim of the present study was to develop a probioticated cucumber juice by optimizing the concentration of prebiotic (inulin), stevia (Stevia rebudiana) and inoculum (Lactobacillus plantarum) by varying one factor at a time. Cucumber juice with 2% prebiotic had a pH 3.76, acidity 0.239%, total sugars 198.2 μg/mL, reducing sugars 102.3 μg/mL and microbial viability of 1.36 × 10⁸ colony forming units (CFU)/mL, respectively. The juice with 3% stevia had a pH 3.59, acidity 0.388%, total sugars 214 μg/mL, reducing sugars 156.7 μg/mL and microbial viability of 6.7 × 10⁷ CFU/mL, respectively. The juice with 3% inoculum size had a pH 3.82, acidity 0.39%, total sugars 128.3 μg/mL, reducing sugars 198.6 μg/mL and microbial viability of 3.5 × 10⁸ CFU/mL, respectively. Maximum growth of the probiotic (L. plantarum) in the cucumber juice was obtained at the end of 48 h of fermentation for 2% prebiotic, 3% stevia and 3% inoculum size without much changes in nutritional and organoleptic properties. The probiotic strain namely L. plantarum proved its ability and suitability to ferment cucumber juice. The growth of the probiotic in the cucumber juice was found maximum with 2% prebiotic, 3% stevia and 3% inoculum size at the end of 48 h of fermentation with minimal changes in nutritional value and organoleptic characteristics with a good shelf life at 4 °C in polyethylene terephthalate bottles.     

DNA Barcodes Distinguish <Emphasis Type="Italic">Withania somnifera</Emphasis> and <Emphasis Type="Italic">Withania ashwagandha</Emphasis>

On the basis of morphology, chemical profiling, crossing ability, amplified fragment length polymorphism and comparison of nucleotide sequences of nuclear ribosomal internal transcribed spacer (ITS), the cultivated form of Withania somnifera, a species of therapeutic value, has been circumscribed as a new species, Withania ashwagandha. The present study was undertaken to ascertain whether the two species can be distinguished on the basis of DNA barcoding. Six barcode loci, ITS, ITS2, matK (maturase K), rbcL (ribulose-bisphosphate carboxylase/oxygenase, large subunit), rpoC1 (RNA polymerase-β′ subunit, main catalytic subunit) and trnH-psbA spacer (transfer RNA for histidine-photosystem II protein D1 spacer) from W. somnifera, W. ashwagandha, their hybrid, and ‘ashwagandha’ market samples were amplified, sequenced, and compared. ITS, ITS2 and matK distinguished two species on the basis of phylogenetic tree method. Likewise, BLAST 1 analysis based on ITS, ITS2, matK, and rbcL individually discriminated two species. However, on the basis of Kimura 2 Parameter distances, two species could not be distinguished as the requirement of a distinct barcode gap—the highest intraspecific distance being lower than the lowest interspecific distance—was not met by any of the loci. If compared by character-based method, ITS, ITS2 and matK sequences of the two species had distinct diagnostic nucleotides (pure character attributes) at nine, four and one positions, respectively. Interestingly, all market samples co-segregated and shared character attributes with W. ashwagandha.     

Rizatriptan <Emphasis Type="Italic">vs.</Emphasis> ibuprofen in migraine: a randomised placebo-controlled trial

The objective of this study was to compare the efficacy of rizatriptan and ibuprofen in migraine. The study was a randomised placebo-controlled trial in a tertiary care teaching hospital. Migraine patients with <8 attacks/months were included. One hundred and fifty-five migraine patients were randomised to rizatriptan 10 mg (53), ibuprofen 400 mg (52) and placebo (50). Efficacy was assessed by headache relief, and headache freedom at 2 h and 24 h. Two-hour headache relief, was noted in 73% in rizatriptan, 53.8% in ibuprofen and 8% in placebo groups. Headache freedom was achieved in 37.7% in rizatriptan, 30.8% in ibuprofen and 2% in placebo groups. Rizatriptan was superior to ibuprofen and placebo in relieving headache at 2 h but not at 24 h. Side effects were noted in 9 patients in rizatriptan, 8 in ibuprofen and 3 in placebo, all of which were nonsignificant. Rizatriptan and ibuprofen are superior to placebo. Rizatriptan is superior to ibuprofen in relieving headache, associated symptoms and functional disability.     

Nutritional Requirements to Improve Delta-Endotoxins Production of <Emphasis Type="Italic">Bacillus</Emphasis><Emphasis Type="Italic">thuringiensis</Emphasis> var. <Emphasis Type="Italic">kurstaki</Emphasis> Using Mixed Designs Modelling

Bacillus thuringiensis kurstaki is a soil bacterium that produces insecticidal toxins called delta-endotoxins. In order to increase the toxic crystal concentrations in a low-cost culture medium and thus improve the biopesticide quality to control insect pests, the Plackett–Burman screening method was applied. It was shown a tool to evaluate the significance of the selected seven factors (KH₂PO₄, K₂HPO₄, MgSO₄, MnSO₄, FeSO₄, soybean meal, starch) which are necessary to the production of the delta-endotoxins. This was performed into two different shake flasks (250 and 500 ml). The main factors that affected the production of delta-endotoxins are shown to be soybean meal, starch, and FeSO₄ in 250 ml culture flasks. In 500 ml culture flasks, soybean meal and FeSO₄ are the principal factors influencing the delta-endotoxin production. The multiple linear regression, a method applied as the merging dataset of the two Plackett–Burman designs, established that soybean meal and starch are the factors positively affecting the production of delta-endotoxins, in contrast to FeSO₄. Furthermore, the available oxygen in culture flasks showed no significant negative effect on delta-endotoxin production. This study revealed that mixed method designs were useful to identify the significance and the effect of hidden culture parameters.     

Larvicidal Activity of <Emphasis Type="Italic">Colocasia esculenta,Eclipta prostrata</Emphasis> and <Emphasis Type="Italic">Wrightia tinctoria</Emphasis> Leaf Extract Against <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis>

Control of mosquitoes by using chemical insecticides creates several problems including development of resistance. This leads to find out alternative methods via plant products. Viewing this in mind, methanolic extracts of Colocasia esculenta, Eclipta prostrata and Wrightia tinctoria leaves were tested against II, III, IV instars and pupa of filarial vector Culex quinquefasciatus. The LC₅₀ value obtained for IV instar larvae of Cx. quinquefasciatus is 165.697 ppm for C. esculenta while it is 114.257 ppm for E. prostrata and 210.298 ppm for W. tinctoria. Of the three plants studied E. prostrata is most effective in controlling mosquito larvae than the others.     

Bioefficacy of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (Balsamo) Vuillemin and <Emphasis Type="Italic">Lecanicillium lecanii</Emphasis> Zimmerman against <Emphasis Type="Italic">Thrips tabaci</Emphasis> Lindeman

Bioefficacy of Beauveria bassiana (Balsamo) Vuillemin and Lecanicillium lecanii Zimmerman in comparison with their commercial formulations along with standard check insecticide, Fenvalerate 20 EC were evaluated against onion thrips, Thrips tabaci Lindeman under greenhouse as well as field conditions. The results revealed that the standard check fenvalerate 20 EC @ 0.0075 % showed significantly the highest cumulative corrected mortality of 97.84 % followed by commercial formulation of B. bassiana, Myco-Jaal @ 1 × 10⁸ spores/mL which showed 80.90 % mortality. The laboratory cultured B. bassiana showed percent mortalities of 74.11, 71.69 and 78.48 % for the concentrations of 1.23 × 10⁷, 1.23 × 10⁶ and 1.23 × 10⁸ spores/mL, respectively. However, these concentrations were statistically at par on all the days of observation. Thrips mortality gradually increased with the increase in concentrations of fungal preparations and days of observations. Similar trend was also observed in L. lecanii experiment. Under field conditions, Fenvalerate 20 EC @ 0.0075 % recorded highest mortality of T. tabaci (90.10 %) followed by commercial formulation of V. lecanii (Phule Bugicide @ 2 × 108 cfu/g) with 74.90 % mortality. All the concentrations of fungal concentrations gave low mortality ranging from 9.40 to 10.10 % and 7.10 to 7.40 % at 2 days after treatment (DAT) of B. bassiana and L. lecanii, respectively. The standard check of Fenvalerate 20 EC @ 0.0075 % was highly toxic and showed significantly maximum percent reduction (90.50 %) of T. tabaci population in both the experiments. The present study clearly shows that these entomopathogens may be integrated with existing integrated pest management (IPM) practices for management of T. tabaci.     

Evaluation of Cultivated and Wild <Emphasis Type="Italic">Allium</Emphasis> Accessions for Resistance to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">cepae</Emphasis>

Fusarium basal rot (FBR) caused by Fusarium oxysporum f. sp. cepae (FOC) is a highly destructive soil borne disease incurring heavy damage in pre and post harvest onion and garlic crops worldwide. Only a few onion lines exhibit partial resistance against the pathogen and there is a need for identification of more effective resistance sources for use in breeding programmes. Selected sets of wild onion and garlic accession and seven related Allium species were screened for resistance to Fusarium basal rot using three FOC isolates. FOC infection revealed significant variation among the evaluated Allium species (at P = 0.001). A. sativum accession ‘CBT-As153’ showed high level of resistance to each isolate while A. cepa accession ‘CBT-Ac77’ exhibited intermediate resistance. Among related Allium species, A. fistulosum, A. roylei and A. schoenoprasum were highly resistant, A. tuberosum had mixed response while A. griffithianum was susceptible. Further, the root density of Allium species negatively correlated with disease incidence for different FOC isolates. Thus, the present study suggests that besides related Allium species, A. sativum ‘CBT-As153’ can be used as a potential donor of FBR resistance for genetic improvement of onion and garlic in India.     

Motion-Artifact-Free <Emphasis Type="Italic">In Vivo</Emphasis> Imaging Utilizing Narcotized Avian Embryos <Emphasis Type="Italic">In Ovo</Emphasis>

Purpose  

The chick embryo in ovo is a well-accessible and economical in vivo model, but its use in molecular imaging has been limited because of motion artifacts on resulting images. The purpose of this study was to develop a method using narcotics to inhibit motility and to perform motion-artifact-free imaging of living chick embryos in ovo.     

Clinical relevance of <Emphasis Type="Italic">Aspergillus</Emphasis> isolation from respiratory tract samples in critically ill patients

Introduction  

The diagnosis of invasive pulmonary aspergillosis, according to the criteria as defined by the European Organisation for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG), is difficult to establish in critically ill patients. The aim of this study is to address the clinical significance of isolation of Aspergillus spp. from lower respiratory tract samples in critically ill patients on the basis of medical and radiological files using an adapted diagnostic algorithm to discriminate proven and probable invasive pulmonary aspergillosis from Aspergillus colonisation.     

Tissue Culture Independent <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> Mediated <Emphasis Type="Italic">In Planta</Emphasis> Transformation Method for Tropical Maize (<Emphasis Type="Italic">Zea mays.L</Emphasis>)

Tropical maize is recalcitrant to tissue culture regeneration because of its poor response to in vitro regeneration after transformation. In this context, the present study has developed the tissue culture independent in planta transformation protocol for tropical maize by transferring plumular meristem cells of germinating seeds of tropical maize genotype through Agrobacteriumtumefaciens infection. The protocol was developed by using Agrobacterium strain EHA105 containing vector pCAMBIA3301 carrying cry1Ab, gus and bar genes. The expression of transgene gus in T₀ plants was confirmed by measuring the hydrolysis rate of the fluorescent substrate 4-methylumbelliferyl-beta-D-glucuronide (MUG) assay whereas the presence of cry1Ab gene was confirmed by polymerase chain reaction and T₀ plants were allowed to grow in glass house into whole plant until maturity and were selfed to produce seeds of T₁ generation. The presence of transgene and its segregation was studied in T₁ generation through Southern and enzyme-linked immunosorbent assay confirming the presence of transgene and its expression respectively. The developed protocol is cost-effective, efficient and genotype independent without involvement of any tissue culture procedure and can generate a relatively large number of transgenic plants in approximately 75–90 days.     

First Attempt of Complete Rearing of Tea Looper, <Emphasis Type="Italic">Biston</Emphasis> (=<Emphasis Type="Italic">Buzura</Emphasis>)<Emphasis Type="Italic"> suppressaria</Emphasis>, on Artificial and Natural Diet

Complete rearing of the looper pest, Biston (=Buzura) suppressaria (Guen.) (Lepidoptera: Geometridae) through generations has been attempted for the first time on artificial diet as an alternative to tea-leaf diet. A comparative study on performance of the pest on its natural host, tea (Camellia sinensis, O’Kuntz), and on the newly formulated artificial diet revealed adequacy and superiority of the latter diet. The development period of B. suppressaria was shorter (48 days) on artificial diet, with a better survival rate than that on tea. On artificial diet the species showed a greater effective rearing rate with production of heavier pupae and adults. Nutritional indices determined for advanced looper stage tilted in favour of artificial diet as compared to that on tea with significantly higher relative growth rate, efficiency of conversion of ingested food, efficiency of conversion of digested food, approximate digestibility, production index and significantly lower maintenance cost.     

Sedation and renal impairment in critically ill patients: a <Emphasis Type="Italic">post hoc</Emphasis> analysis of a randomized trial

Introduction  

Not sedating critically ill patients reduces the time patients receive mechanical ventilation, decreases the time in the intensive care department and reduces the total hospital length of stay. We hypothesized that no sedation improves hemodynamic stability, decreases the need for vasoactive drugs, diminishes the need for extra fluids and lowers the risk of acute kidney injury.     

Recurrent <Emphasis Type="Italic">Clostridium difficile</Emphasis> colitis

Clostridium difficile is the most common cause of nosocomial infectious diarrhea that is usually treated adequately with standard treatment of metronidazole or vancomycin. Relapse or recurrent infection can occur in certain patients and this can be very difficult to treat. The authors have stated that they do not have a significant financial interest or other relationship with any product manufacturer or provider of services discussed in this article. The authors also do not discuss the use of off-label products, which includes unlabeled, unapproved, or investigative products or devices.     

Ultrasonic-enhanced gentamicin transport through colony biofilms of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis>

The hypothesis that ultrasound increases antibiotic transport through biofilms of Escherichia coli and Pseudomonas aeruginosa was investigated using colony biofilms. Biofilms grown on membrane filters were transferred to nutrient agar containing 50µg/ml gentamicin. A smaller filter was placed on top of the biofilm and a blank concentration disk was situated atop the filter. Diffusion of antibiotic through the biofilms was allowed for 15, 30, or 45min at 37°C. Some of these biofilms were exposed to 70-kHz ultrasound and others were not. Each concentration disk was then placed on a nutrient agar plate spread with a lawn of E. coli. The resulting zone of inhibition was used to calculate the amount of gentamicin that was transported through the biofilm into the disk. The E. coli and P. aeruginosa biofilms grown for 13 and 24h were exposed to two different ultrasonic power densities. Ultrasonication significantly increased the transport of gentamicin through the biofilm. Insonation of biofilms of E. coli for 45min more than doubled the amount of gentamicin compared to their noninsonated counterparts. For P. aeruginosa biofilms, no detectable gentamicin penetrated the biofilm within 45min without ultrasound; however, when insonated (1.5W/cm²) for 45min, the disks collected more than 0.45µg antibiotic. Ultrasonication significantly increased transport of gentamicin across biofilms that normally blocked or slowed gentamicin transport when not exposed to ultrasound. This enhanced transport may be partially responsible for the increased killing of biofilm bacteria exposed to combinations of antibiotic and ultrasound.     

Molecular Authentication of Medicinal Plant, <Emphasis Type="Italic">Swertia</Emphasis><Emphasis Type="Italic">chirayita</Emphasis> and its Adulterant Species

Swertia is ethno-medicinally an important genus belonging to family Gentianaceae. Swertia chirayita is used as imperative medicinal plant in Indian system of medicine. However, this species has been frequently adulterated due to its high demand and scarcity. Authentication of this species was needed to protect consumers and conservation measures and to find out the alternative source. Deoxyribose nucleic acid (DNA) extracted from fifteen samples of six species belonging to different localities, were used as templates. Four candidate barcodes were amplified by polymerase chain reaction and sequence analysis was executed by Z-Big Dye Terminator Cycle Sequencing v.3. Sequenced products were analyzed on automated applied biosystems 3730XI analyzer. Identification was performed by using molecular evolutionary genetics analysis 5 software (version 5.1). The amplification efficiency of all DNA barcodes [megakaryocyte-associated tyrosine kinase (matK), ribulose-bisphosphate carboxylase (rbcL), photosystem II protein D1- stuctural RNA- His tRNA (psbA-trnH) and internal transcribed spacer region (ITS)] was 100 %. Here, the highest interspecific divergence provided by ITS is 11.87 % and intraspecific by psbA-trnH being 10.22 % as compared to matK (5.04 %) and rbcL (0.99 %). ITS region proves robust molecular marker for differing the S. chirayita from its related adulterant species. All barcoding regions indicate that S. chirayita and S. minor both are more closely related than other Swertia species. Findings showed that DNA barcoding is an efficient tool for identification and authentication of S. chirayita. Use of S. minor as substitute to S. chirayita can be advocated.