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.     

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.     

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.     

<Emphasis Type="Italic">Leuconostoc</Emphasis> bacteremia in three patients with malignancies

Leuconostoc is a Gram-positive coccus characterized by its resistance to glycopeptide antibiotics. Generally, this bacterium is susceptible to β-lactam antibiotics; however, here we present a leukemia patient who developed leuconostoc bacteremia during antimicrobial therapy with carbapenem. The appropriate choice of antibiotics at optimal doses enables leuconostoc infection to be overcome, even in compromised hosts. We report 3 cases of leuconostoc bacteremia: the leukemia case which was successfully treated, along with discussions of two other cases with malignancies.     

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.     

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.     

Investigating Disease Controlling Ability of <Emphasis Type="Italic">Brassica</Emphasis> Volatiles and Their Compatibility with <Emphasis Type="Italic">Trichoderma harzianum</Emphasis>

The present investigation aimed to test the in vitro toxicity of Brassica volatiles against soil-borne plant pathogens (Rhizoctonia solani Kuhn, Sclerotium rolfsii Sacc., Fusarium oxysporum f.sp. ciceris (Padwick) Matuo & K. Sato and Sclerotinia sclerotiorum (Lib.) de Bary) and biocontrol fungus Trichoderma harzianum Rifai. Moreover, the response of T. harzianum to toxic volatiles was also studied in terms of glucanase and chitinase gene expression. A strong fungistatic effect of B. alba treatment was recorded against all the test pathogens. Noticeably, S. sclerotiorum manifested least sensitivity among all the tested pathogens. All T. harzianum isolates were less sensitive as compared to the assayed pathogens. Based on the in vitro study involving 6 T. harzianum isolates, T₅₅ and Th-R showed least, while T₃₉ showed highest sensitivity to the volatiles. Interestingly, the patterns relating to the effect of volatiles on inoculum density of T. harzianum isolates were similar to their effect on fungus growth in vitro. More importantly, expression of chitinase and glucanase genes in different Trichoderma isolates was up-regulated, which could improve the biocontrol activity of T. harzianum. Therefore, here the authors envisage that combining biocontrol and biofumigation has the potential to provide sustainable and cost-effective strategies to manage soil-borne plant pathogens.     

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.     

No regional spread of vancomycin-resistant enterococci with <Emphasis Type="Italic">vanA</Emphasis> or <Emphasis Type="Italic">vanB</Emphasis> in Kitakyushu,Japan

Outbreaks of vancomycin-resistant enterococci (VRE) infection occur sporadically in Japan, and their frequency has been gradually increasing. We experienced a nosocomial outbreak of VRE in two hospitals in the city of Kitakyushu, and the spread of VRE strains was suspected in this area. To examine the prevalence rate of infection and colonization of VRE in Kitakyushu, we screened a total of 24297 clinical samples from patients in hospitals and clinics in Kitakyushu from October through December 2002 for VRE. The isolates screened as positive for VRE accounted for 2.3% (566/24297) of the tested clinical samples. Polymerase chain reaction (PCR) analyses for vanA, vanB, vanC1, and vanC2/3 were performed to confirm the screening test results. Neither vanA nor vanB genes were detected in any isolates. The 265 vanC1-positive isolates were Enterococcus gallinarum, and the 150 vanC2/3-positive isolates were E. casseliflavus. Other Enterococcus species were negative in this PCR-detection test. In this study, the PCR procedure was considered reliable and successful because although neither vanA nor vanB was detected, vanC1 and vanC2/3 were completely detectable. Therefore, we concluded that the regional spread of VRE with vanA and vanB had not occurred in Kitakyushu in 2002. In the near future, the prevalence of VRE with vanA or vanB is likely to increase in Japan, as it has in other countries. We should continue to find and prevent nosocomial outbreaks of infection and colonization by VRE.     

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.     

<Emphasis Type="Italic">In vitro</Emphasis> and <Emphasis Type="Italic">in vivo</Emphasis> evaluation of a new active heat moisture exchanger

Introduction  

In order to improve the efficiency of heat moisture exchangers (HMEs), new hybrid humidifiers (active HMEs) that add water and heat to HMEs have been developed. In this study we evaluated the efficiency, both in vitro and in vivo, of a new active HME (the Performer; StarMed, Mirandola, Italy) as compared with that of existing HMEs (Hygroster and Hygrobac; Mallinckrodt, Mirandola, Italy).     

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.     

Pulmonary nocardiosis caused by <Emphasis Type="Italic">Nocardia exalbida</Emphasis> complicating <Emphasis Type="Italic">Pneumocystis</Emphasis> pneumonia in an HIV-infected patient

A 47-year-old man with optimally controlled type-2 diabetes mellitus and chronic hepatitis B was admitted to a local hospital because of a 1-week history of cough and high-grade fever. He was diagnosed with Pneumocystis pneumonia (PCP) and Klebsiella pneumonia from a chest radiograph and sputum. Simultaneously, he was found to have HIV infection with a CD4 count of 76/μl. Despite alteration of treatment secondary to the development of allergic reaction to trimethoprim–sulfamethoxazole (TMP–SMX), the patient was able to complete a 3-week therapy for PCP after being switched to pentamidine isetionate. After the treatment of PCP, he was referred to our hospital for the initiation of anti-HIV therapy. He presented with recurrent high-grade fever of a few days’ duration prior to his initial visit, which subsequently led to his admission. Chest computed tomography (CT) showed the enlargement of a previously identified infiltrate in the left upper lung field, and the sputum culture upon admission was positive for Gram-positive branching rods; the organism was later identified as Nocardia exalbida. Due to his allergy to sulfonamide, the patient was treated with imipenem (IMP) and amikacin (AMK) given intravenously for 17 days, followed by garenoxacin (GRNX) taken orally for 6 months, without any adverse effects. The chest infiltrate resolved completely, and he remains stable without relapse 8 months after the completion of the therapy. Pulmonary nocardiosis should be considered as a differential diagnosis of recurring pneumonia in immunocompromised patients, especially in HIV-infected individuals. Oral administration of GRNX following IMP and AMK can be used as an alternative to TMP–SMX therapy in cases of pulmonary nocardiosis caused by N. exalbida.