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Fas‐associated death domain protein (FADD) is an apoptotic adapter molecule that recruits caspases and facilitates the formation of death‐inducing signaling complexes in mammals. In lower animals, FADD forms a complex with immune deficiency and death‐related ced‐3/Nedd2‐like caspase to elicit antimicrobial responses. In this study, we characterized the FADD sequence of Tenebrio molitor (TmFADD) using molecular informatics to understand its role in immune surveillance of the host against microorganisms. The 618 bp open reading frame of TmFADD encodes a polypeptide of 204 amino acids. The protein includes two promiscuous domains, the death‐effector domain and the death domain, found at the N‐ and C‐termini, respectively. TmFADD is clustered with the coleopteran FADD sequences in the phylogenetic tree. Homology‐based structural analysis showed an α‐helical model with a QMEAN score of 0.58. The expression of TmFADD mRNA was conspicuous in the larval and early pupal stages. TmFADD mRNA was distributed in the immune tissues of the larva and was found predominately in the gut of the adult. Infection of the host with Candida albicans, Escherichia coli or Staphylococcus aureus increased the expression of TmFADD mRNA, suggesting that these microorganisms trigger the innate immunity of the host. The silencing of TmFADD transcripts resulted in an increased susceptibility of T. molitor larvae to infection with E. coli, but not to infection with C. albicans or S. aureus. Hence, these results indicate that FADD plays a role in the defense of T. molitor and could open avenues for understanding insect immunity. [ABSTRACT FROM AUTHOR]

Edible insects such as the mealworm (Tenebrio molitor) are emerging as an alternative animal protein resource to meet the needs of a growing global population. However, despite the successful mass production of edible insects, research into practical compound feed for edible insect is still insufficient. In this study, mealworms were fed wheat bran, layer chicken feed mash, chicken pellets and cattle pellets for 1 to 36 days, after which their body length was measured. In the groups fed with chicken feed mash, chicken pellets or cattle pellets, the mean body lengths were 2.88, 2.92 and 2.95 cm, respectively, similar to the 2.95 cm of the control T. molitor fed with wheat bran. Individual weight measurements yielded similar results: control, 7.87 g; chicken feed mash, 7.43 g; chicken pellets, 7.60 g; and cattle pellets, 7.63 g. Analysis of livestock feed intake showed that the intake was 290.4 g in control insects, 137.7 g in insects fed chicken feed mash, 220.2 g in insects fed chicken pellets and 230.3 g in insects fed cattle pellets. In the excretion analysis, the excretion of the control insects (161.60 g) was larger than that of the insects fed with chicken feed mash (93.37 g), chicken pellets (93.40 g) or cattle pellets (133.87 g). An analysis of the nutritional content of T. molitor based on nine major nutrients showed no significant differences between the control and the test groups. These results can be used to improve the price competitiveness and quality of T. molitor for commercial development. [ABSTRACT FROM AUTHOR]

Selective herbicides are used to control undesirable vegetation or weeds in fields without harming crops. Herbicide use for weed management can directly impact the densities of insect pests in agricultural communities as a result of insect mortality during and immediately after application. In insects, the glutathione S‐transferase (GST) enzyme is involved in both the detoxification process and the defense of cellular membranes against oxidative damage. In this study, two TmGSTs (TmGST‐iso1 and TmGST‐iso2) were identified and characterized to elucidate the GST family in Tenebrio molitor. Among the developmental stages of T. molitor, eggs had the highest expression levels of TmGST‐iso1. TmGST‐iso2 expression was highest in the pre‐pupal stage. TmGST‐iso1 expression was high in the guts of early and late larvae, whereas TmGST‐iso2 expression was not observed in early larvae. Adults, both male and female, had the highest levels of TmGST‐iso1 mRNA expression in the gut and reproductive organs, and the highest levels of TmGST‐iso2 expression in the reproductive organs. Quantitative polymerase chain reaction was used to investigate the impact of treatment with butachlor on the mRNA expression of TmGST‐iso1 and TmGST‐iso2 in larvae. TmGST‐iso1 expression increased in the butachlor‐treated group after 3 and 24 h, whereas TmGST‐iso2 expression peaked at 24 h after treatment. This study provides vital information about the detoxifying activities of T. molitor. [ABSTRACT FROM AUTHOR]

In insects, the production of antimicrobial peptides is considered to be the solo arm of the innate immune response. The Toll and immune deficiency pathways are two major signaling pathways that lead to the production of antimicrobial peptides as final effectors. The dynamic functions of Toll/Toll‐like receptors have been thoroughly reviewed in mammals and Drosophila. During the last decade, we have attempted to clarify the immunological roles of different Toll receptor variants and their ligands in Tenebrio molitor. Accordingly, we showed that TmToll‐8, −9, and −10 mRNA transcripts are induced following systemic injections of Gram‐negative, Gram‐positive, and fungal infections. Our data revealed harmonic expression patterns of TmToll‐8, −9, and −10 throughout the developmental stages of healthy individuals and in response to infections in the examined tissues (Malpighian tubules, gut, fat bodies, and hemolymph), illustrating similar evolutionary conversions of different Toll receptor variants in T. molitor. Taken together, our findings highlight the immunological actions of TmToll‐8, −9, and −10 in response to pathogenic insults in T. molitor. [ABSTRACT FROM AUTHOR]

Agricultural intensification has led to significant increases in production, but the overuse of pesticides and associated hazards pose threats to biodiversity and ecological functions. Catalase (CAT), a key antioxidant enzyme, plays a crucial role in alleviating oxidative stress by directly interacting with toxins. In this study, we identified three CAT isoforms in Tenebrio molitor (TmCAT‐iso1, TmCAT‐iso2 and TmCAT‐iso3). These CATs possess a CAT domain, tetramer interface sites and a heme‐binding pocket. We examined the expression of Tm catalases across all developmental stages and in specific tissues using quantitative real time polymerase chain reaction (qRT‐PCR) experiments. Our findings demonstrate that TmCAT‐iso1 and TmCAT‐iso3 exhibit peak expression in young and late larval stages, respectively, whereas TmCAT‐iso2 shows peak expression during the egg and pre‐pupal stages. Tissue distribution analysis revealed the high expression of TmCAT‐iso1 and TmCAT‐iso2 in larval hemocytes, whereas TmCAT‐iso3 is predominantly expressed in larval Malpighian tubules. Furthermore, injection with chlorantraniliprole significantly elevated mRNA expression levels of TmCAT‐iso1, TmCAT‐iso2, and TmCAT‐iso3 in larval groups, compared with control groups. Our study highlights the distinct developmental stages and tissues where TmCATs are expressed. We also elucidated the effects of pesticide application on the expression of each TmCAT, revealing the physiological characteristics of CATs in response to these pesticides, which are dose‐ and time‐dependent in T. molitor. [ABSTRACT FROM AUTHOR]

Mealworms beetles, Tenebrio molitor, are the limelight next-generation food for humans due to their high nutrient contents. Since Tenebrio molitor is used as feed for pets and livestock in addition to their ability to decompose polystyrene and plastic waste, it is recognized as an insect with an industrial core value. Therefore, it is important to study the immune mechanism related to the development and infection of mealworms for mass breeding purposes. The immune deficiency (Imd) signaling is one of the main pathways with pivotal roles in the production of antimicrobial peptides (AMPs). Transforming growth factor-β activated kinase (TAK1) is one of the Imd pathway components, forms a complex with TAK1 binding protein 2 (TAB2) to ultimately help activate the transcription factor Relish and eventually induce host to produce AMPs. Relatively, little has been revealed about TAK1 in insect models, especially in the T. molitor. Therefore, this study was conducted to elucidate the function of TmTak1 in T. molitor. Our results showed that the highest and lowest mRNA expression of TmTak1 were found in egg and young larvae respectively. The tissue-specific expression patterns were reported in the gut of T. molitor larvae and the fat bodies of adults. Systemic microbial challenge illustrated TmTak1 high expression following the fungal infection in all dissected tissues except for the whole body. However, silencing TmTak1 experiments showed that the survivability of T. molitor larvae affected significantly following Escherichia coli infection. Accordingly, AMP induction after TmTak1 knock down was mainly reported in the integument and the fat bodies. [ABSTRACT FROM AUTHOR]

Although Toll-like receptors have been widely identified and functionally characterized in mammalian models and Drosophila, the immunological function of these receptors in other insects remains unclear. Here, we explored the relevant innate immune response of Tenebrio molitor (T. molitor) Toll-3 against Gram-negative bacteria, Gram-positive bacteria, and fungal infections. Our findings indicated that TmToll-3 expression was mainly induced by Candida albicans infections in the fat bodies, gut, Malpighian tubules, and hemolymph of young T. molitor larvae. Surprisingly, Escherichia coli systemic infection caused mortality after TmToll-3 knockdown via RNA interference (RNAi) injection, which was not observed in the control group. Further analyses indicated that in the absence of TmToll-3, the final effector of the Toll signaling pathway, antimicrobial peptide (AMP) genes and relevant transcription factors were significantly downregulated after E. coli challenge. Our results indicated that the expression of almost all AMP genes was suppressed in silenced individuals, whereas the expression of relevant genes was positively regulated after fungal injection. Therefore, this study revealed the immunological involvement of TmToll-3 in T. molitor in response to systematic infections. [ABSTRACT FROM AUTHOR]

The antimicrobial roles of Toll-like receptors have been mainly identified in mammalian models and Drosophila. However, its immunological function in other insects has yet to be fully clarified. Here, we determined the innate immune response involvement of TmToll-2 encountering Gram-negative, Gram-positive, and fungal infection. Our data revealed that TmToll-2 expression could be induced by Escherichia coli, Staphylococcus aureus, and Candida albicans infections in the fat bodies, gut, Malpighian tubules, and hemolymph of Tenebrio molitor young larvae. However, TmToll-2 silencing via RNAi technology revealed that sole E. coli systemic infection caused mortality in the double-strand RNA TmToll-2-injected group compared with that in the control group. Further investigation indicated that in the absence of TmToll-2, the final effector of Toll signaling pathway, antimicrobial peptide (AMP) genes and relevant transcription factors were significantly downregulated, mainly E. coli post-insult. We showed that the expression of all AMP genes was suppressed in the main immune organ of insects, namely, fat bodies, in silenced individuals, while the relevant expressions were not affected after fungal infection. Thus, our research revealed the immunological roles of TmToll-2 in different organs of T. molitor in response to pathogenic insults. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) are considered to be candidate effectors for eliciting humoral immune responses against infectious pathogens in the host. Cecropins are α‐helical peptides of 30–40 amino acids, which are known to permeabilize bacterial membranes, and play authoritative roles in the innate immune system of insects. In the present study, we identified the full‐length open reading frame (ORF) encoding the Tenebrio molitor cecropin‐2 (TmCec2) gene using the Tribolium castaneum cecropin‐2 (TcCec2) gene to query a T. molitor Rnaseq database. Phylogenetic analysis revealed close identity of TmCec2 with TcCec2. TmCec2 was ubiquitously expressed in the insect during all developmental stages, with the highest expression observed in the adult. Tissue‐specific TmCec2 expression was highest in larval hemocytes and in the adult integument and hemocytes. Microbial challenge experiments revealed that TmCec2 was highly induced in response to gram‐positive and gram‐negative bacteria, and fungi. These data provide credible evidence for a putative role of TmCec2 in insect innate immunity against a plethora of pathogens. [ABSTRACT FROM AUTHOR]

Defensins are a major family of antimicrobial peptides that serve as the innate immune defense of both vertebrates and invertebrates. Due to their antimicrobial, chemotactic, and regulatory activities, Defensins have been exploited for their therapeutic potential. Insect Defensins are cysteine‐rich and contain an N‐terminal loop, α‐helix, and antiparallel β‐sheet, forming a "cysteine‐stabilized alpha beta (CSαβ)" or "loop–helix‐sheet" structure. In this study, we identified the full‐length open reading frame (ORF) sequences of Defensin (TmDef) and Defensin‐like (TmDef‐like) genes from the mealworm beetle Tenebrio molitor using in silico methods. TmDef and TmDef‐like genes encode the peptides of 72 and 71 amino acid residues, respectively. TmDefensin is comprised of a Defensin domain and the TmDefensin‐like is comprised of a signal peptide of 21 amino acid residues. Phylogenetic analysis revealed close similarities of TmDefensin with the Defensin of Acalolepta luxuriosa of the longhorn beetle family. The expression of TmDef mRNA was found to be greater than that of TmDef‐like mRNA and was mostly expressed in the pupal and adult stages. Tissue distribution showed high expression of TmDef‐like mRNA in larval hemocytes, gut, integument, and fat body, while in adults, the expression was high in gut and hemocytes. Following bacterial and fungal stimulation in vivo, TmDef was upregulated at 24 h post‐infection in whole body, fat body, and hemocytes of the larvae. Even TmDef‐like mRNA was upregulated in the gut and hemocytes at 12 and 9 h post‐infection respectively. These results suggest that TmDef and TmDef‐like genes play important roles in protecting T. molitor from microbial contact. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) are effector candidates that elicit humoral immunity in invertebrates. AMPs facilitate bacterial clearance by either physically disrupting the microbial membranes or the intracellular targets. In the Coleopteran pest, Tenebrio molitor, transcriptional regulation of AMPs has been studied in the context of innate immune signaling cascades and antimicrobial immunity. Here, we report the transcriptional response of three AMP genes, Coleoptericin A, B, and C (TmCole A, B and C) in T. molitor in response to bacterial (Escherichia coli, Staphylococcus aureus), and fungal (Candida albicans) challenges. These genes were expressed essentially in the gut and hemocytes followed by the integument tissue of the T. molitor larva. Further, these genes were highly expressed in the late‐larval, pupal, and early adult stages. Furthermore, while all of these transcripts were highly upregulated in the fat body and Malpighian tubules after bacterial challenge, TmCole B and TmCole C were induced in the gut after E. coli challenge. Fungal stimulation was not required for the upregulation of the transcription of Coleoptericin genes in T. molitor. [ABSTRACT FROM AUTHOR]

Toll and IMD pathways regulate antimicrobial innate immune responses in insect model systems. The transcriptional activation of antimicrobial peptides (AMPs) confers humoral immunity in the host against invaded pathogens. The IKK kinase complex (IKKα, IKKβ, and the regulatory subunit IKKγ/NEMO) centrally regulates the NF-κB response to various stimuli. It triggers an appropriate antimicrobial immune response in the host. In this study, a Tm IKKβ (or Tm Ird5) homolog was screened from the RNA-seq database of the coleopteran beetle, Tenebrio molitor. A single exon characterizes the Tm IKKβ gene, and the open reading frame (ORF) comprises of 2112 bp that putatively encodes a polypeptide of 703 amino acid residues. Tm IKKβ contains a serine/threonine kinase domain and is phylogenetically close to Tribolium castaneum IKKβ homolog (Tc IKKβ). TmIKKβ transcripts were highly expressed in the early pupal (P1) and adult (A5) stages. Among the tissues, TmIKKβ showed higher expression in the integument of the last instar larvae and the fat body and hemocytes of 5-day-old adults. TmIKKβ mRNA was upregulated post- E. coli challenge to the host. Moreover, RNAi-based TmIKKβ mRNA silencing increased host larvae' susceptibility against E. coli, S. aureus and C. albicans. TmIKKβ RNAi in the fat body led to a downregulation in mRNA expression of ten out of fourteen AMP genes, including TmTenecin1, -2, and -4 ; TmDefensin, and -like ; TmColeoptericinA, and -B ; and TmAttacin1a, - 1b, and -2 , suggesting the requirement of the gene in antimicrobial innate immune responses. Further, a decrease in the mRNA expression of NF-κB factors such as TmRelish, TmDorsal1 , and TmDorsal2 in the fat body of T. molitor larvae was observed post-microorganisms challenge. Thus, Tm IKKβ regulates antimicrobial innate immune responses in T. molitor. • We have newly identified IκB kinase β gene from the Tenebrio molitor , named TmIKKβ and domain analysis indicated that it contains one Serine/Threonine protein kinase catalytic domain, suggesting that TmIKKβ may regulate cell signaling by phosphorylation. • The temporal and spatial expression patterns of TmIKKβ transcripts and its induction patterns in responses to microbial challenges indicated that the TmIKKβ may have important roles in development as well as immune responses against microbial challenges. • We have investigated the effects of TmIKKβ RNAi on larval survivability, and expression patterns of antimicrobial peptide (AMP) genes and transcription factors for Toll and Imd pathways to understand the critical function of TmIKKβ in antimicrobial immune responses. [ABSTRACT FROM AUTHOR]

Five genes encoding PhaP family proteins and one phaR gene have been identified in the genome of Burkholderia symbiont strain RPE75. PhaP proteins function as the surface proteins of polyhydroxyalkanoate (PHA) granules, and the PhaR protein acts as a negative regulator of PhaP biosynthesis. Recently, we characterized one phaP gene to understand the molecular cross talk between Riptortus insects and Burkholderia gut symbionts. In this study, we constructed four other phaP gene-depleted mutants (ΔphaP1, ΔphaP2, ΔphaP3, and ΔphaP4 mutants), one phaR gene-depleted mutant, and a phaR-complemented mutant (ΔphaR/phaR mutant). To address the biological roles of four phaP family genes and the phaR gene during insect-gut symbiont interaction, these Burkholderia mutants were fed to the secondinstar nymphs, and colonization ability and fitness parameters were examined. In vitro, the ΔphaP3 and ΔphaR mutants cannot make a PHA granule normally in a stressful environment. Furthermore, the ΔphaR mutation decreased the colonization ability in the host midgut and negatively affected the host insect's fitness compared with wild-type Burkholderia-infected insects. However, other phaP family genedepleted mutants colonized well in the midgut of the fifth-instar nymph insects. However, in the case of females, the colonization rate of the ΔphaP3 mutant was decreased and the host's fitness parameters were decreased compared with the wild-type-infected host, suggesting that the environment of the female midgut may be more hostile than that of the male midgut. These results demonstrate that PhaR plays an important role in the biosynthesis of PHA granules and that it is signifi- cantly related to the colonization of the Burkholderia gut symbiont in the host insects' midgut. Bacterial polyhydroxyalkanoate (PHA) biosynthesis is a complex process requiring several enzymes. The biological roles of PHA granule synthesis enzymes and the surface proteins of PHA granules during host-gut symbiont interactions are not fully understood. Here, we report the effects on colonization ability in the host midguts and the fitness of host insects after feeding Burkholderia mutant cells (four phaP-depleted mutants and one phaR-depleted mutant) to the host insects. Analyses of both synthesized PHA granule amounts and CFU numbers suggest that the phaR gene is closely related to synthesis of the PHA granule and the colonization of the Burkholderia gut symbiont in the host insect's midgut. Like our previous report, this study also supports the idea that the environment of the host midgut may not be favorable to symbiotic Burkholderia cells and that PHA granules may be required to adapt in the host midgut. [ABSTRACT FROM AUTHOR]

Riptortus pedestris , a common pest in soybean fields, harbors a symbiont Burkholderia in a specialized posterior midgut region of insects. Every generation of second nymphs acquires new Burkholderia cells from the environment. We compared in vitro cultured Burkholderia with newly in vivo colonized Burkholderia in the host midgut using biochemical approaches. The bacterial cell envelope of in vitro cultured and in vivo Burkholderia differed in structure, as in vivo bacteria lacked lipopolysaccharide (LPS) O-antigen. The LPS O-antigen deficient bacteria had a reduced colonization rate in the host midgut compared with that of the wild-type Burkholderia . To determine why LPS O-antigen-deficient bacteria are less able to colonize the host midgut, we examined in vitro survival rates of three LPS O-antigen-deficient Burkholderia mutants and lysates of five different midgut regions. The LPS O-antigen-deficient mutants were highly susceptible when cultured with the lysate of a specific first midgut region (M1), indicating that the M1 lysate contains unidentified substance(s) capable of killing LPS O-antigen-deficient mutants. We identified a 17 kDa protein from the M1 lysate, which was enriched in the active fractions. The N-terminal sequence of the protein was determined to be a soybean Kunitz-type trypsin inhibitor. These data suggest that the 17 kDa protein, which was originated from a main soybean source of the R. pedestris host, has antibacterial activity against the LPS O-antigen deficient (rough-type) Burkholderia. [ABSTRACT FROM AUTHOR]

The Riptortus-Burkholderia symbiotic system represents a promising experimental model to study the molecular mechanisms involved in insect-bacterium symbiosis due to the availability of genetically manipulated Burkholderia symbiont. Using transposon mutagenesis screening, we found a symbiosis-deficient mutant that was able to colonize the host insect but failed to induce normal development of host's symbiotic organ. The disrupted gene was identified as purL involved in purine biosynthesis. In vitro growth impairment of the purL mutant and its growth dependency on adenine and adenosine confirmed the functional disruption of the purine synthesis gene. The purL mutant also showed defects in biofilm formation, and this defect was not rescued by supplementation of purine derivatives. When inoculated to host insects, the purL mutant was initially able to colonize the symbiotic organ but failed to attain a normal infection density. The low level of infection density of the purL mutant attenuated the development of the host's symbiotic organ at early instar stages and reduced the host's fitness throughout the nymphal stages. Another symbiont mutant-deficient in a purine biosynthesis gene, purM, showed phenotypes similar to those of the purL mutant both in vitro and in vivo, confirming that the purL phenotypes are due to disrupted purine biosynthesis. These results demonstrate that the purine biosynthesis genes of the Burkholderia symbiont are critical for the successful accommodation of symbiont within the host, thereby facilitating the development of the host's symbiotic organ and enhancing the host's fitness values. [ABSTRACT FROM AUTHOR]

Observational studies on transfusion in trauma comparing high versus low plasma:erythrocyte ratio were prone to survivor bias because plasma administration typically started later than erythrocytes. Therefore, early deaths were categorized in the low plasma:erythrocyte group, whereas early survivors had a higher chance of receiving a higher ratio. When early deaths were excluded, however, a bias against higher ratio can be created. Survivor bias could be reduced by performing before-and-after studies or treating the plasma:erythrocyte ratio as a time-dependent covariate.We reviewed 26 studies on blood ratios in trauma. Fifteen of the studies were survivor bias-unlikely or biased against higher ratio; among them, 10 showed an association between higher ratio and improved survival, and five did not. Eleven studies that were judged survivor bias-prone favoring higher ratio suggested that a higher ratio was superior.Without randomized controlled trials controlling for survivor bias, the current available evidence supporting higher plasma:erythrocyte resuscitation is inconclusive. [ABSTRACT FROM AUTHOR]

We compared cold-pain responses among male current opioid users with and without concurrent benzodiazepine use, long-term ex-users, and healthy controls. Forty-eight current opioid users (14 concurrently using benzodiazepines), 34 ex-users (abstinent for >=1y) and 63 controls received cold-pressor tests. Pain threshold (first reporting pain) and pain tolerance (total immersion time) were recorded. Pain thresholds were similar in ex-users and current users; pain tolerance was similar in ex-users and controls. Net pain tolerance (endurance) in ex-users was intermediate between the other two groups. Current users showed higher pain threshold and shorter pain tolerance than controls (p<0.05). Current users not co-using benzodiazepines showed the lowest pain tolerance and net pain tolerance, and differed significantly from controls, ex-users, and current users co-using benzodiazepines (p<0.05). Neuroticism was higher in current users than in the other two groups (p<0.001), extraversion marginally lower (p<0.05); net pain tolerance differences remained significant after controlling for these. Benzodiazepine use modulates pain tolerance in opioid users. Pain responses altered by opioid use may partially recover with abstinence. [ABSTRACT FROM AUTHOR]

Surgical emphysema is a well-recognised complication of laparoscopic surgery, but its impact on end-tidal carbon dioxide levels and carbon dioxide elimination is seldom reported and may not be fully appreciated by anaesthetists. Four cases are presented where extensive surgical emphysema occurred during laparoscopic surgery. The visual display of the anaesthetic record using the software program Monitor showed substantial rises in end-tidal carbon dioxide levels and allowed calculation of the carbon dioxide elimination, which increased two- to three-fold above normal levels. Having a visual record of carbon dioxide changes facilitated the recognition of surgical emphysema in three out of the four cases. Strategies such as estimating and tracking changes in carbon dioxide elimination from the minute ventilation and end-tidal carbon dioxide levels may assist in early identification, and palpating for surgical emphysema is recommended during laparoscopy if other causes of increased carbon dioxide levels are excluded. [ABSTRACT FROM AUTHOR]

We describe a novel technique, previously applied to small children, for adult one-lung anaesthesia in which a single-lumen endotracheal tube is used with an endobronchial balloon blocker The main aims of the technique are to reduce the likelihood of cephalad displacement of the balloon into the trachea and to facilitate directional placement of the endobronchial balloon. We present five illustrative cases of one-lung anaesthesia in patients of adult size, in which the endotracheal tube-endobronchial balloon technique was considered preferable to the use of a double-lumen tube technique. The situations included difficult intubation, need for postoperative ventilation, a tortuous trachea and an unexpected need to perform one-lung anaesthesia. The technique involved deliberate placement of the endotracheal tube tip near the carina to block cephalad dislodgement of the blocker The chance of the balloon blocking the endotracheal tube tip could be further reduced by having the intraluminal endobronchial balloon blocker emerge through the Murphy eye. [ABSTRACT FROM AUTHOR]

Objective: To evaluate, by systematic review, the efficacy of heliox on respiratory mechanics and outcomes in patients with acute asthma.Methods: The search strategy included searching electronic databases (MEDLINE, EMBASE, and The Cochrane Library) and the references of relevant articles. Study quality was assessed based on allocation concealment. Randomized controlled trials (RCTs) comparing heliox to an air-oxygen mixture (airO(2)) as an adjunct treatment in patients with acute asthmatic attacks were analyzed. For the qualitative portion of the analysis, all reports of the use of heliox in patients with acute asthma were included.Results: Four RCTs (n = 278) were found to have a common respiratory parameter (peak expiratory flow rate as a percentage of predicted) suitable for meta-analysis. Within the 92% confidence interval (CI), there was a small benefit with the use of heliox compared to airO(2) (weighted mean difference, + 3%; 95% CI, - 2 to + 8%). There was also a slight improvement in the dyspnea index (weighted mean difference, 0.60; 95% CI, 0.04 to 1.16) with the use of heliox over airO(2). Overall, five RCTs, one nonrandomized unblinded parallel trial, one retrospective case-matched control trial, three case series, and one case report had results in favor of heliox; one RCT and one case series showed no improvement with heliox; one RCT showed a possible detrimental effect with heliox; and 1 small RCT was inconclusive. Most investigators did not prevent entrainment of room air during heliox use or compensate for the lower nebulizing efficiency of heliox.Conclusion: Based on surrogate markers, heliox may offer mild-to-moderate benefits in patients with acute asthma within the first hour of use, but its advantages become less apparent beyond 1 h, as most conventionally treated patients improve to similar levels, with or without it. The effect of heliox may be more pronounced in more severe cases. There are insufficient data on whether heliox can avert tracheal intubation, or change intensive care and hospital admission rates and duration, or mortality. [ABSTRACT FROM AUTHOR]

The cystine knot protein Spätzle is a Toll receptor ligand that modulates the intracellular signaling cascade involved in the nuclear factor kappa B (NF-κB)-mediated regulation of antimicrobial peptide (AMP)-encoding genes. Spätzle-mediated activation of the Toll pathway is critical for the innate immune responses of insects against Gram-positive bacteria and fungi. In this study, the open reading frame (ORF) sequence of Spätzle-like from T. molitor (TmSpz-like) identified from the RNA sequencing dataset was cloned and sequenced. The 885-bp TmSpz-like ORF encoded a polypeptide of 294 amino acid residues. TmSpz-like comprised a cystine knot domain with six conserved cysteine residues that formed three disulfide bonds. Additionally, TmSpz-like exhibited the highest amino acid sequence similarity with T. castaneum Spätzle (TcSpz). In the phylogenetic tree, TmSpz-like and TcSpz were located within a single cluster. The expression of TmSpz-like was upregulated in the Malpighian tubules and gut tissues of T. molitor. Additionally, the expression of TmSpz-like in the whole body and gut of the larvae was upregulated at 24 h post-E. coli infection. The results of RNA interference experiments revealed that TmSpz-like is critical for the viability of E. coli-infected T. molitor larvae. Eleven AMP-encoding genes were downregulated in the E. coli-infected TmSpz-like knockdown larvae, which suggested that TmSpz-like positively regulated these genes. Additionally, the NF-κB-encoding genes (TmDorX1, TmDorX2, and TmRelish) were downregulated in the E. coli-infected TmSpz-like knockdown larvae. Thus, TmSpz-like plays a critical role in the regulation of AMP production in T. molitor in response to E. coli infection. [ABSTRACT FROM AUTHOR]

A letter to the editor about reducing finger cuts among anesthetists when opening glass ampoules is presented.

Recently, our group demonstrated that the bean bug, Riptortus pedestris , is a good experimental symbiosis model to study the molecular cross-talk between the host insect and the gut symbiont. The Burkholderia symbiont is orally acquired by host nymphs from the environment in every generation. However, it is still unclear how Riptortus specifically interacts with entomopathogens that are abundant in the environmental soil. In preliminary experiments, we observed that a potent entomopathogen, Serratia marcescens , can colonize the midgut of Riptortus insects and was recovered from the midgut when Serratia cells were orally administered, suggesting that this pathogenic bacterium can escape host immune defenses in the salivary fluid. We examined how orally fed Serratia cells can survive in the presence of antimicrobial substances of the Riptortus salivary fluid. In this study, a 15 kDa trialysin-like protein from the salivary gland of R. pedestris and a potent virulence factor of Serratia cells, a serralysin metalloprotease, from the culture medium of S. marcescens were successfully purified to homogeneity. When the purified Riptortus trialysin (rip-trialysin) was incubated with purified serralysin, rip-trialysin was specifically hydrolyzed by serralysin, leading to the loss of antimicrobial activity. These results clearly demonstrated that a potent virulent metalloprotease of S. marcescens functions as a key player in the escape from the salivary fluid-mediated host immune response, resulting in successful colonization of S. marcescens in the host midgut. [ABSTRACT FROM AUTHOR]

Valuable insect models have tremendously contributed to our understanding of innate immunity and symbiosis. Bean bug, Riptortus pedestris , is a useful insect symbiosis model due to harboring cultivable monospecific gut symbiont, genus Burkholderia . Bean bug is a hemimetabolous insect whose immunity is not well-understood. However, we recently identified three major antimicrobial peptides of Riptortus and examined the relationship between gut symbiosis and host immunity. We found that the presence of Burkholderia gut symbiont positively affects Riptortus immunity. From studying host regulation mechanisms of symbiont population, we revealed that the symbiotic Burkholderia cells are much more susceptible to Riptortus immune responses than the cultured cells. We further elucidated that the immune-susceptibility of the Burkholderia gut symbionts is due to the drastic change of bacterial cell envelope. Finally, we show that the immune-susceptible Burkholderia symbionts are able to prosper in host owing to the suppression of immune responses of the symbiotic midgut. [ABSTRACT FROM AUTHOR]

We generated a Burkholderia mutant, which is deficient of an N -acetylmuramyl- l -alanine amidase, AmiC, involved in peptidoglycan degradation. When non-motile Δ amiC mutant Burkholderia cells harboring chain form were orally administered to Riptortus insects, Δ amiC mutant cells were unable to establish symbiotic association. But, Δ amiC mutant complemented with amiC gene restored in vivo symbiotic association. Δ amiC mutant cultured in minimal medium restored their motility with single-celled morphology. When Δ amiC mutant cells harboring single-celled morphology were administered to the host insect, this mutant established normal symbiotic association, suggesting that bacterial motility is essential for the successful symbiosis between host insect and Burkholderia symbiont. [ABSTRACT FROM AUTHOR]

Many insects possess symbiotic bacteria that affect the biology of the host. The level of the symbiont population in the host is a pivotal factor that modulates the biological outcome of the symbiotic association. Hence, the symbiont population should be maintained at a proper level by the host's control mechanisms. Several mechanisms for controlling intracellular symbionts of insects have been reported, while mechanisms for controlling extracellular gut symbionts of insects are poorly understood. The bean bug Riptortus pedestris harbors a betaproteobacterial extracellular symbiont of the genus Burkholderia in the midgut symbiotic organ designated the M4 region. We found that the M4B region, which is directly connected to the M4 region, also harbors Burkholderia symbiont cells, but the symbionts therein are mostly dead. A series of experiments demonstrated that the M4B region exhibits antimicrobial activity, and the antimicrobial activity is specifically potent against the Burkholderia symbiont but not the cultured Burkholderia and other bacteria. The antimicrobial activity of the M4B region was detected in symbiotic host insects, reaching its highest point at the fifth instar, but not in aposymbiotic host insects, which suggests the possibility of symbiont- mediated induction of the antimicrobial activity. This antimicrobial activity was not associated with upregulation of antimicrobial peptides of the host. Based on these results, we propose that the M4B region is a specialized gut region of R. pedestris that plays a critical role in controlling the population of the Burkholderia gut symbiont. The molecular basis of the antimicrobial activity is of great interest and deserves future study. [ABSTRACT FROM AUTHOR]

The beneficial effects of Acanthopanax divaricatus var. albeofructus (ADA) extracts have been assessed by mutagenic and anti-mutagenic activities by Ames test. Mutation of Salmonella typhimurium strains TA 98, TA 100, TA1535, TA1537, and Escherichia coli WP2 uvr A was assayed in duplicates by the procedure of Maron and Ames in the presence or absence of S9 mix. As a result, ADA extracts were not mutagenic for S. typhimurium strains TA 98, TA 100, TA1535, TA1537, and E. coli by the Ames assay. Anti-mutagenic activity was assayed by the Ames mutagenicity assay using histidine mutant of S. typhimurium strains TA 98 and TA 100, using the plate-incorporation method. 2-Aminoanthrancene (2-AA), 2-(2-furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2), and sodium azide (NaN3) were used as the mutagens. ADA extracts showed a strong anti-mutagenic activity against 2-AA-induced mutagenesis which requires liver-metabolizing enzymes, and the same extract exhibited inhibitory effects on AF-2 and NaN3-induced mutagenesis in the absence of liver-metabolizing enzymes. The data indicate that ADA extracts contain anti-mutagenic activities against typical mutagens. The anti-mutagenic property of ADA provides additional health supplemental value to the other claimed therapeutic properties of the plant. [ABSTRACT FROM AUTHOR]

Vertebrates rely on the most sophisticated adaptive immunity to defend themselves against various pathogens. This includes immunologic memory cells, which mount a stronger and more effective immune response against an antigen after its first encounter. Unlike vertebrates, invertebrates' defense completely depends on the innate immunity mechanisms including humoral and cell-mediated immunity. Furthermore, the invertebrate equivalent of the memory cells was discovered only recently. Since the discovery of transgenerational immune priming (TGIP) in crustaceans, numerous findings have proven the IP in invertebrate classes such as insects. TGIP can be induced through maternal priming pathways such as transcriptional regulation of antimicrobial peptides, and also paternal IP including the induction of proPO system activity. We appraise the diversity and specificity of IP agents to provide sustained immunologic memory in insects, particularly T. molitor in the review. An understanding of IP (more so TGIP) response in T. molitor will deepen our knowledge of invertebrate immunity, and boost the mass-rearing industry by reducing pathogen infection rates. • Unlike mammals, solo arm of defense mechanism in invertebrates is innate immunity, particularly, antimicrobial peptide production. • The ability of being immunized against a preexisting elicitor, was a matter of uncertainty until a wile ago. • This phenomenon known as "immune priming", can be transferred to progenies by both parents. • The diversity and specificity of immune priming in insects more precisely, Tenebrio molitor has been investigated in numbers of studies. • Understanding of immune priming in insects can provide us with novel strategies and applications at the field during T. molitor mass rearing. [ABSTRACT FROM AUTHOR]

To investigate the relationship between personality, club-drug use and high-risk drug-related behaviour, 360 club-drug users and 303 non-drug users in Hong Kong were assessed on measures of two impulsivity dimensions, reward drive and rash impulsivity, and a related trait of punishment sensitivity. The most frequently used drugs were ketamine, ecstasy, and cannabis, with the majority of participants using two or more drugs on any one occasion. Club-drug users were more rash-impulsive and reward-driven, and less punishment-sensitive than non-drug users (p<0.001). Rash impulsivity, but not reward drive or punishment sensitivity, was significantly (p<0.001) associated with risky drug-related behaviour. There was no association between any personality traits and preferred drug. These findings suggest that, while those who use club drugs are generally more impulsive and less punishment-sensitive, some discrete facets of impulsivity are associated with differing patterns of drug-use behaviour. [ABSTRACT FROM AUTHOR]

Recent studies have provided molecular evidence that gut symbiotic bacteria modulate host insect development, fitness and reproduction. However, the molecular mechanisms through which gut symbionts regulate these aspects of host physiology remain elusive. To address these questions, we prepared two different Riptortus-Burkholderia insect models, Burkholderia gut symbiont-colonized (Sym) Riptortus pedestris insects and gut symbiont-noncolonized (Apo) insects. Upon LC-MS analyses, juvenile hormone III skipped bisepoxide (JHSB 3) was newly identified from Riptortus Apo- and Sym-female and male adults' insect hemolymph and JHSB 3 titer in the Apo- and Sym-female insects were measured because JH is important for regulating reproduction in adult insects. The JHSB 3 titer in the Sym-females were consistently higher compared to those of Apo-females. Since previous studies reported that Riptortus hexamerin-α and vitellogenin proteins were upregulated by the topical abdominal application of a JH-analog, chemically synthesized JHSB 3 was administered to Apo-females. As expected, the hexamerin-α and vitellogenin proteins were dramatically increased in the hemolymph of JHSB 3 -treated Apo-females, resulting in increased egg production compared to that in Sym-females. Taken together, these results demonstrate that colonization of Burkholderia gut symbiont in the host insect stimulates biosynthesis of the heteroptera-specific JHSB 3 , leading to larger number of eggs produced and enhanced fitness in Riptortus host insects. • Heteroptera-specific JHSB 3 is a bona fide juvenile hormone in R. pedestris insect. • The Burkholderia gut symbiont modulates titter of JHSB 3 in Riptortus female insect. • The expression of hemolymph proteins are induced by topical application of JHSB 3. • This study describes molecular relationship between gut-symbiont and host juvenile hormone. [ABSTRACT FROM AUTHOR]

Recent studies have suggested that gut symbionts modulate insect development and reproduction. However, the mechanisms by which gut symbionts modulate host physiologies and the molecules involved in these changes are unclear. To address these questions, we prepared three different groups of the insect Riptortus pedestris : Burkholderia gut symbiont-colonized (Sym) insects, Burkholderia -non-colonized (Apo) insects, and Burkholderia -depleted (Sym Burk- ) insects, which were fed tetracycline. When the hemolymph proteins of three insects were analyzed by SDS-PAGE, the hexamerin-α, hexamerin-β and vitellogenin-1 proteins of Sym-adults were highly expressed compared to those of Apo- and Sym Burk- -insects. To investigate the expression patterns of these three genes during insect development, we measured the transcriptional levels of these genes. The hexamerin-β gene was specifically expressed at all nymphal stages, and its expression was detected 4–5 days earlier in Sym-insect nymphs than that in Apo- and Sym Burk- -insects. However, the hexamerin-α and vitellogenin-1 genes were only expressed in adult females, and they were also detected 6–7 days earlier and were 2-fold higher in Sym-adult females than those in the other insects. Depletion of hexamerin-β by RNA interference in 2nd instar Sym-nymphs delayed adult emergence, whereas hexamerin-α and vitellogenin-1 RNA interference in 5th instar nymphs caused loss of color of the eggs of Sym-insects. These results demonstrate that the Burkholderia gut symbiont modulates host development and egg production by regulating production of these three hemolymph storage proteins. [ABSTRACT FROM AUTHOR]

The relation between gut symbiosis and immunity has been reported in various animal model studies. Here, we corroborate the effect of gut symbiont to host immunity using the bean bug model. The bean bug, Riptortus pedestris , is a useful gut symbiosis model due to the monospecific gut symbiont, genus Burkholderia . To examine the effect of gut symbiosis to host immunity, we generated the gut symbiont-harboring (symbiotic) insect line and the gut symbiont-lacking (aposymbiotic) insect line. Upon bacterial challenges, the symbiotic Riptortus exhibited better survival than aposymbiotic Riptortus . When cellular immunity was inhibited, the symbiotic Riptortus still survived better than aposymbioic Riptortus , suggesting stronger humoral immunity. The molecular basis of the strong humoral immunity was further confirmed by the increase of hemolymph antimicrobial activity and antimicrobial peptide expression in the symbiotic insects. Taken together, our data clearly demonstrate that Burkhoderia gut symbiont positively affect the Riptortus systemic immunity. [ABSTRACT FROM AUTHOR]

OBJECTIVE: Intraabdominal hypertension reduces organ blood flow. Restoring abdominal perfusion pressure (APP) may restore renal blood flow, especially when sepsis is present. The effects of intra-abdominal pressure (IAP), followed by restoration of APP with norepinephrine, on renal blood flow were determined. DESIGN: Longitudinal study with bacteremia after nonbacteremic (control) conditions. SETTING: University animal laboratory. SUBJECTS: Ten anesthetized mongrel dogs. INTERVENTIONS: IAP was raised to 10, 20, and 30 mm Hg, using intra-abdominal bags filled with saline. After each intervention, decompression was achieved by emptying the bag. Bacteremia was induced by injection of Escherichia coli. Cardiac output and renal blood flow were measured using surgically placed flow probes. Norepinephrine infusion was used to restore the mean arterial pressure to baseline at each IAP. A hypervolemic circulation was maintained throughout by infusing saline. MEASUREMENTS AND MAIN RESULTS: Induction of bacteremia resulted in significant decreases in blood pressure, cardiac output, and renal blood flow (p < .01). Serial increases in IAP decreased cardiac output and renal blood flow both in control and bacteremic dogs (p < .001). These decreases were substantially corrected by abdominal decompression. In nonbacteremic control conditions, restoring APP back to baseline with norepinephrine did not fully restore cardiac output and renal blood flow (p < .001). However, in bacteremic conditions, norepinephrine was able to substantially restore cardiac output and renal blood flow to bacteremic baseline at all levels of IAP. In bacteremic conditions, the renal perfusion fraction returned to bacteremic baseline levels after correction of APP with norepinephrine and after decompression. CONCLUSIONS: Restoration of APP using norepinephrine improves renal blood flow in bacteremic animals with IAPs up to 30 mm Hg, and maintaining a therapeutic APP may preserve renal blood flow in patients with intra-abdominal hypertension who are at risk of IAP-induced renal injury but who have yet to meet accepted criteria for surgical decompression. [ABSTRACT FROM AUTHOR]

IKKγ/NEMO is the regulatory subunit of the IκB kinase (IKK) complex, which regulates the NF-κB signaling pathway. Within the IKK complex, IKKγ/NEMO is the non-catalytic subunit, whereas IKKα and IKKβ are the structurally related catalytic subunits. In this study, TmIKKγ was screened from the Tenebrio molitor RNA-Seq database and functionally characterized using RNAi screening for its role in regulating T. molitor antimicrobial peptide (AMP) genes after microbial challenges. The TmIKKγ transcript is 1521 bp that putatively encodes a polypeptide of 506 amino acid residues. TmIKKγ contains a NF-κB essential modulator (NEMO) and a leucine zipper domain of coiled coil region 2 (LZCC2). A phylogenetic analysis confirmed its homology to the red flour beetle, Tribolium castaneum IKKγ (TcIKKγ). The expression of TmIKKγ mRNA showed that it might function in diverse tissues of the insect, with a higher expression in the hemocytes and the fat body of the late-instar larvae. TmIKKγ mRNA expression was induced by Escherichia coli, Staphylococcus aureus, and Candida albicans challenges in the whole larvae and in tissues such as the hemocytes, gut and fat body. The knockdown of TmIKKγ mRNA significantly reduced the survival of the larvae after microbial challenges. Furthermore, we investigated the tissue-specific induction patterns of fourteen T. molitor AMP genes in TmIKKγ mRNA-silenced individuals after microbial challenges. In general, the mRNA expression of TmTenecin1, -2, and -4; TmDefensin1 and -2; TmColeoptericin1 and 2; and TmAttacin1a, 1b, and 2 were found to be downregulated in the hemocytes, gut, and fat body tissues in the TmIKKγ-silenced individuals after microbial challenges. Under similar conditions, TmRelish (NF-κB transcription factor) mRNA was also found to be downregulated. Thus, TmIKKγ is an important factor in the antimicrobial innate immune response of T. molitor. [ABSTRACT FROM AUTHOR]

Autophagy is an important process by which pathogens and damaged or unused organelles are eliminated. The role of autophagy in development and the immune response to pathogens is well established. Autophagy-related protein 8 (Atg8) is involved in the formation of the autophagosome and, with the help of the serine protease Atg4, mediates the delivery of both vesicles and the autophagosome to the vacuole. Here, we cloned the Aedes albopictus autophagy-related protein 8 (AaAtg8) gene and characterized its role in the innate immunity of the mosquito against microbial infections. AaAtg8 is comprised of an open reading frame (ORF) region of 357 bp encoding a polypeptide of 118 amino acid residues. A domain analysis of AaAtg8 revealed an Atg8 ubiquitin-like domain, Atg7/Atg4 interaction sites, and peptide binding sites. The AaAtg8 mRNA expression was high in the Malpighian tubules and heads of both sugar-fed and blood-fed adult female mosquitoes. The expression level of AaAtg8 mRNA increased in the midgut and abdominal carcass following being challenged with Listeria monocytogenes. To investigate the role of AaAtg8 in the innate immune responses of Ae. albopictus, AaAtg8 gene-silenced adult mosquitoes were challenged by injection or by being fed microorganisms in blood. High mortality rates were observed in mosquitoes in which AaAtg8 was silenced after challenges of microorganisms to the host by blood feeding. This suggests that Atg8-autophagy plays a critical role in the gut immunity in Ae. albopictus. [ABSTRACT FROM AUTHOR]