Erratum to "The influence of ecological factors on cnidarian venoms" [Toxicon: X 9-10C (2021) 100067].

[This corrects the article DOI: 10.1016/j.toxcx.2021.100067.].

Inducing metamorphosis in the irukandji jellyfish Carukia barnesi.

Here we utilize chemical ecology as a tool to manipulate the biological system of a small, but highly venomous to humans, cubozoan jellyfish, Carukia barnesi. We trialled a range of chemical reagents including indole compounds, 9-cis-retinoic acid and lugols solution to induce metamorphosis between the polyp and medusa life stages. An optimum method was determined resulting in a 90% metamorphosis rate to healthy medusa by exposing the polyps to 1 µM of 5-methoxy-2-methylindole for 24 h. Of note is that chemical exposure time significantly impacts health and metamorphosis rates in this species. We also present a theoretical mechanism for the chemical/biological interactions occurring during metamorphosis. This is a significant methodological advancement which now enables rearing of this animal en mass in aquaria-a world first for this species-which will subsequently supply and facilitate venom research into this understudied jellyfish.

The influence of ecological factors on cnidarian venoms.

Venom research is often focussed on medical relevance, novel compounds and venom evolution, whilst studying the relationship between a venom and its environment - venom ecology - has been conducted to a lesser extent. Given the projected environmental changes envisioned to occur with global warming, it is pertinent now more than ever, to highlight this topic. Here we review literature examining the influence of ecological factors such as environmental temperature, salinity, ontogeny, geographic location and diet on cnidarian venoms. This review provides an exclusive focus on the cnidarian phylum and encompasses all available published, peer-reviewed literature to our knowledge regarding the ecological factors influencing venom. We find a startling lack of research into the effects of both environmental and biological factors on venoms, with very few to no studies available per category. Importantly, research does exist that suggest these ecological processes may influence other marine or terrestrial venoms, thus we recommend future research is needed to explore this concept in cnidarians.

Altered mucosa-associated microbiota in the ileum and colon of neonatal calves in response to delayed first colostrum feeding.

The present study investigated whether delaying the first feeding of colostrum affected ileum and colon mucosa-associated microbiota in calves. Twenty-seven male Holstein calves were randomly assigned to 1 of 3 groups, fed colostrum at 45 min, 6 h, and 12 h after birth, respectively. Ileum and colon mucosa were collected at 51 h after birth, and their associated microbial profiles were assessed using amplicon sequencing. Both ileum and colon mucosa-associated microbiota were predominated by genus Escherichia-Shigella. The negative correlation between the molar proportion of short-chain fatty acids (SCFA) and ileum mucosa-associated opportunistic pathogens, and the positive correlation between the molar proportion of SCFA and colon mucosa-associated beneficial bacteria, suggest that SCFA might play an important role in maintaining the gut health of 2-d-old calves. A higher relative abundance of ileum mucosa-associated Enterococcus and Streptococcus was detected when the first colostrum feeding was delayed for 12 h. The relative abundance of colon mucosa-associated Lactobacillus tended to be lower in calves fed colostrum 12 h than those under the other 2 treatments, whereas that of Faecalibacterium tended to be lower in calves fed colostrum immediately after birth than those fed colostrum 6 and 12 h after birth, respectively. Our findings suggest that delayed first colostrum feeding affects the establishment of ileum and colon mucosa-associated bacteria, which may have long-term effects on gut health of calves.

Effects of dry period length on metabolic status, fertility, udder health, and colostrum production in 2 cow breeds.

Shortening the dry period improves postpartum energy balance, which has potential positive effects on metabolic health. This concept has been mainly studied in Holstein cows. The aim of this study was to assess the effect of a short dry period of 4 wk, compared with a standard dry period of 8 wk, on the metabolic status, progesterone profiles, health, and colostrum quality of dairy cows of 2 breeds, Swedish Red (SR) and Swedish Holstein (SH), not treated with antibiotics at dry off. The IgG uptake in calves was also studied to reflect the colostrum quality when shortening the dry period. Cows of both SH and SR were blocked by breed and parity and then randomly allocated to a short dry period of 4 wk (4W, n = 43) or a conventional dry period of 8 wk (8W, n = 34). Blood samples were collected wk -8, -4, -2, -1 and 1, 2, 3, 4, 6, 9, and 12 relative to calving. Prepartum, cows with a 4-wk dry period had higher concentrations of nonesterified fatty acids and lower concentrations of insulin-like growth factor-1 and insulin than 8W cows. Postpartum, plasma concentration of nonesterified fatty acids was lower, whereas plasma insulin and insulin-like growth factor-1 tended to be higher for 4W cows than for 8W cows. Plasma concentration of ß-hydroxybutyrate did not differ between dry period lengths. Swedish Holstein cows with a 4W dry period responded with a lower concentration of insulin prepartum than SR and SH on an 8W dry period. The dry period length had no effect on the proportion of disturbed progesterone profiles; disturbed progesterone profiles occurred in 30% of the 4W cows and 47% of the 8W cows. In this trial, only 48.8% of the SR cows had a normal progesterone profile, which differed from the SH where 76.5% had a normal profile. Fertility-related diseases (endometritis, pyometra, anestrus, ovarian cyst) did not differ between the 2 dry period groups: 21% in the 8W group versus 12% in the 4W group, whereas mastitis tended to be more common: 26% of the 4W cows versus 9% of the 8W cows. A short dry period resulted in less colostrum but with a higher content of protein and somatic cell count. Calves were fed colostrum from their dam, and the IgG and total protein in plasma did not differ between calves to mothers with different a dry period length. Shortening the dry period could improve metabolic status in cows of both SH and SR breed postpartum, without compromising the colostrum quality. Health and progesterone profiles were not affected by the dry period length for SH or SR in this study.

Effects of dry period length on milk production and energy balance in two cow breeds.

Shortening the dry period (DP) has been proposed as a strategy to improve energy balance (EB) in cows in early lactation. This study evaluated the effects of shortening the DP on milk yield (MY), EB and residual feed intake (RFI) in two breeds; Swedish Red (SR) and Swedish Holstein (SH). Cows were blocked by breed and parity and then randomly assigned to one of two treatments; short DP of 4 weeks (4W, n=43) or conventional DP of 8 weeks (8W, n=34). Cows were kept and fed under the same conditions, except for the 4 weeks when the 4W group were still lactating prepartum and thus kept with the lactating cows. Milk yield and BW were recorded and body condition score (BCS) was rated from 10 weeks prepartum to 12 weeks postpartum. Dry matter intake (DMI) was recorded for lactating cows postpartum. Milk yield was reduced by 6.75 kg/day during the first 12 weeks postpartum (P<0.001) for the 4W cows compared with 8W cows, but there was no significant difference in total MY (3724 kg compared with 3684 kg, P=0.7) when the milk produced prepartum was included. Protein content was higher in 4W cows (3.42%) than in 8W cows (3.27%) (P<0.001) postpartum. In the 8W group, cows lost more BCS after calving (P<0.05). Cows of SR breed had higher BCS than cows of SH breed (SR=3.7, SH=3.2, P<0.001), but no differences in BW were found between breed and treatment. Energy balance was improved for cows in the 4W group (P<0.001), while feed efficiency, expressed as RFI, was reduced for 4W cows than for 8W cows (5.91 compared with -5.39, P<0.01). Shortening the DP resulted in improved EB postpartum with no difference between the breeds and no milk losses when including the milk produced prepartum.

Celecoxib reduces narcotic use and pain following augmentation mammaplasty.

BACKGROUND: Narcotics have traditionally been used to control pain after augmentation mammaplasty, but they have adverse side effects, including addiction potential, clouded sensorium, nausea, and respiratory depression. Alternative strategies for managing postoperative pain are expensive, cumbersome, and also have their own risks. While long-term use of celecoxib has been associated with an increased risk of serious adverse cardiovascular effects, no problems have been reported with short-term use. OBJECTIVE: The purpose of this study was to determine whether the addition of celecoxib, a selective nonsteroidal anti-inflammatory, to an analgesic regimen reduced narcotic use and pain following augmentation mammaplasty. METHODS: One hundred patients underwent submuscular augmentation mammaplasty with smooth saline-filled mammary prostheses using an intravenous sedation technique. Group A (N = 50) used hydrocodone to manage postoperative pain. Group B (N = 50) used celecoxib 400 mg 1 to 2 hours before surgery and then daily in addition to hydrocodone postoperatively. Narcotic use, incidence of nausea, and complications were recorded. Pain was assessed daily with a Likert pain scale from 0 (no pain) to 10 (severe pain). RESULTS: Group A used 110 +/- 34 mg hydrocodone during the immediate 7-day postoperative period and reported an average pain scale score of 5.1. Group B, which used celecoxib, used 34 +/- 22 mg hydrocodone during the same period with an average pain scale score of 3.3. These differences were statistically significant (P < .05). Group B experienced 53% less nausea than Group A. There were no significant differences between the groups regarding age, implant size, or complications. CONCLUSIONS: Perioperative celecoxib administration in patients undergoing augmentation mammaplasty significantly reduced postoperative narcotic use, pain, and nausea. Its use should facilitate the patient's ability to resume everyday activities following surgery.

The interferon-induced double-stranded RNA-activated protein kinase PKR will phosphorylate serine, threonine, or tyrosine at residue 51 in eukaryotic initiation factor 2alpha.

The family of eukaryotic initiation factor 2alpha (eIF2alpha) protein kinases plays an important role in regulating cellular protein synthesis under stress conditions. The mammalian kinases PKR and HRI and the yeast kinase GCN2 specifically phosphorylate Ser-51 on the alpha subunit of the translation initiation factor eIF2. By using an in vivo assay in yeast, the substrate specificity of these three eIF2alpha kinases was examined by substituting Ser-51 in eIF2alpha with Thr or Tyr. In yeast, phosphorylation of eIF2 inhibits general translation but derepresses translation of the GCN4 mRNA. All three kinases phosphorylated Thr in place of Ser-51 and were able to regulate general and GCN4-specific translation. In addition, both PKR and HRI were found to phosphorylate eIF2alpha-S51Y and stimulate GCN4 expression. Isoelectric focusing analysis of eIF2alpha followed by detection using anti-eIF2alpha and anti-phosphotyrosine-specific antibodies demonstrated that PKR and HRI phosphorylated eIF2alpha-S51Y on Tyr in vivo. These results provide new insights into the substrate recognition properties of the eIF2alpha kinases, and they are intriguing considering the potential for alternate substrates for PKR in cellular signaling and growth control pathways.

Escherichia coli mrsC is an allele of hflB, encoding a membrane-associated ATPase and protease that is required for mRNA decay.

The mrsC gene of Escherichia coli is required for mRNA turnover and cell growth, and strains containing the temperature-sensitive mrsC505 allele have longer half-lives than wild-type controls for total pulse-labeled and individual mRNAs (L. L. Granger et al., J. Bacteriol. 180:1920-1928, 1998). The cloned mrsC gene contains a long open reading frame beginning at an initiator UUG codon, confirmed by N-terminal amino acid sequencing, encoding a 70,996-Da protein with a consensus ATP-binding domain. mrsC is identical to the independently identified ftsH gene except for three additional amino acids at the N terminus (T. Tomoyasu et al., J. Bacteriol. 175:1344-1351, 1993). The purified protein had a Km of 28 microM for ATP and a Vmax of 21.2 nmol/microg/min. An amino-terminal glutathione S-transferase-MrsC fusion protein retained ATPase activity but was not biologically active. A glutamic acid replacement of the highly conserved lysine within the ATP-binding motif (mrsC201) abolished the complementation of the mrsC505 mutation, confirming that the ATPase activity is required for MrsC function in vivo. In addition, the mrsC505 allele conferred a temperature-sensitive HflB phenotype, while the hflB29 mutation promoted mRNA stability at both 30 and 44 degrees C, suggesting that the inviability associated with the mrsC505 allele is not related to the defect in mRNA decay. The data presented provide the first direct evidence for the involvement of a membrane-bound protein in mRNA decay in E. coli.

The Escherichia coli mrsC gene is required for cell growth and mRNA decay.

We have identified a gene in Escherichia coli that is required for both the normal decay of mRNA and RNA synthesis. Originally designated mrsC (mRNA stability), the mrsC505 mutation described here is, in fact, an allele of the hflB/ftsH locus (R.-F. Wang et al., J. Bacteriol. 180:1929-1938, 1998). Strains carrying the thermosensitive mrsC505 allele stopped growing soon after the temperature was shifted to 44 degrees C but remained viable for several hours. Net RNA synthesis stopped within 20 min after the shift, while DNA and protein synthesis continued for over 60 min. At 44 degrees C, the half-life of total pulse-labeled RNA rose from 2.9 min in a wild-type strain to 5.9 min in the mrsC505 single mutant. In an rne-1 mrsC505 double mutant, the average half-life was 19.8 min. Inactivating mrsC significantly increased the half-lives of the trxA, cat, secG, and kan mRNAs, particularly in an mrsC505 pnp-7 rnb-500 rne-1 multiple mutant. In addition, Northern analysis showed dramatic stabilizations of full-length mRNAs in a variety of mrsC505 multiple mutants at 44 degrees C. These results suggest that MrsC, directly or indirectly, controls endonucleolytic processing of mRNAs that may be independent of the RNase E-PNPase-RhlB multiprotein complex.

Targeted ribozymes reveal a conserved function of the Drosophila paired gene in sensory organ development.

The Drosophila paired (prd) gene, the founding member of the PAX gene family, is required for normal embryonic segmentation and is re-expressed later in development in the head and developing CNS. As for most embryonically active genes, global defects resulting from loss of early prd function obscure an analysis of the role of later expression phases. We used inducible targeted ribozymes to functionally 'knock-out' prd at late stages. When prd protein levels in the head are reduced in this fashion, the maxillary chemosensory ventral organs fail to develop and dorsal-lateral cirri rows are disrupted. These studies reveal a role for prd in sensory organ development that appears to be conserved in PAX genes throughout the animal kingdom.

Polyadenylylation helps regulate mRNA decay in Escherichia coli.

As part of our genetic analysis of mRNA decay in Escherichia coli K-12, we examined the effect of the pcnB gene [encoding poly(A) polymerase I] on message stability. Eliminating poly(A) polymerase I (delta pcnB) dramatically stabilized the lpp, ompA, and trxA transcripts. The half-lives of individual mRNAs were increased in both a delta pcnB single mutant and a delta pcnB pnp-7 rnb-500 rne-1 multiple mutant. We also found mRNA decay intermediates in delta pcnB mutants that were not detected in control strains. By end-labeling total E. coli RNA with [32P]pCp and T4 RNA ligase and then digesting the RNA with RNase A and T1, we showed that many RNAs in a wild-type strain contained poly(A) tails ranging from 10 nt to > 50 nt long. When polynucleotide phosphorylase, RNase II, and RNase E were absent, the length (> 100 nt) and number (10- to 20-fold) of the poly(A) tails increased. After transcription initiation was stopped with rifampicin, polyadenylylation apparently continued. Deleting the structural gene for poly(A) polymerase I (pcnB) reduced the amount of 3'-terminal poly(A) sequences by > 90%. We propose a model for the role of polyadenylylation in mRNA decay.

Distal-less is a downstream gene of Deformed required for ventral maxillary identity.

The homeotic selector (HOM) proteins are required for the diversification of the anterior-posterior axis of the Drosophila body plan, assigning unique identities to regional domains of cells comprising one or a few parasegments or segments. The HOM proteins apparently accomplish this task by the transcriptional regulation of numerous downstream genes. At present few downstream genes are known, so models of how downstream genes mediate HOM functions are based more on intuition than information. Our results indicate that Distal-less is a downstream gene of the HOM gene Deformed, and Distal-less function is required for the elaboration of a subset of the maxillary epidermal identities specified by Deformed. The regulatory effect of Deformed on Distal-less is mediated by a ventral maxillary-specific enhancer located 3' of the Distal-less transcription unit. We propose that Deformed and Distal-less, both of which encode homeodomain transcription factors that are persistently expressed in ventral maxillary cells, combinatorially specify a subsegmental code required for a group of cells to differentiate maxillary cirri.

Molecular characterization of the Aspergillus nidulans yA locus.

We investigated the molecular organization of the region of Aspergillus nidulans chromosome I containing yA, a gene encoding the developmentally regulated enzyme conidial laccase. DNA fragments were identified that complemented the yA2 mutation and were shown to correspond to yA by genetic mapping and gene disruption experiments. The molecular map of the region was oriented to the genetic map by testing DNA fragments for their ability to complement a mutation in the tightly linked adE gene. The yA gene codes for a 2200 nucleotide mRNA that is present at low levels in vegetative cells and mature conidia, but accumulates to high levels in sporulating cultures. yA mRNA appears shortly after differentiation of sporogenous phialide cells. It accumulates in two developmentally abnormal mutant strains that produce phialides but is absent from two mutant strains that do not produce phialides. Thus, yA transcription is probably restricted to phialides. This result is discussed in relationship to the physiological roles played by phialides in spore differentiation.

Enrichment for Hydrogen-Oxidizing Acinetobacter spp. in the Rhizosphere of Hydrogen-Evolving Soybean Root Nodules.

Field soybean plants were inoculated with Hup wild-type or H(2) uptake-negative (Hup) mutants of Bradyrhizobium japonicum. For two consecutive summers we found an enrichment for acinetobacters associated with the surfaces of the H(2)-evolving nodules. Soybean root nodules that evolved H(2) had up to 12 times more Acinetobacter spp. bacteria associated with their surfaces than did nodules incapable of evolving H(2). All of the newly isolated strains identified as Acinetobacter obtained from the surfaces of root nodules, as well as known established Acinetobacter strains, were capable of oxidizing H(2), a property not previously described for this alkane-degrading soil bacterium.

Staged infrainguinal revascularization: initial prosthetic above-knee bypass followed by a distal vein bypass for recurrent ischemia. A valid concept for extending limb salvage?

Optimal infrainguinal revascularization should provide limb salvage for the longest duration of time. It is not known whether limb salvage is longer with an initial below-knee popliteal or tibial in situ saphenous vein graft or with staged bypasses; that is, an initial above-knee popliteal prosthetic bypass if feasible, followed by a more distal vein graft should the above-knee prosthetic graft fail. A retrospective review of 197 lower extremity vascular reconstructions performed since 1976 utilizing polytetrafluoroethylene (PTFE), umbilical vein, or in situ saphenous vein was completed. The data were analyzed for differences in limb salvage and prevention of limb threatening ischemia among three subgroups: above-knee prosthetic bypass, below-knee or tibial in situ saphenous vein bypass, and staged reconstructions (above-knee prosthetic bypass with subsequent in situ bypass). The groups were similar with respect to severity of limb threatening ischemia as indicated by mean preoperative ankle-brachial indices. Cumulative secondary limb salvage at 36 months was 73 percent for prosthetic grafts in the above-knee position, 78 percent for in situ saphenous vein grafts in the below-knee or tibial position, and 87 percent for staged reconstruction with an initial prosthetic graft to the above-knee position followed by a distal in situ vein bypass when the prosthetic graft fails.

Venous stasis ulceration. Effectiveness of subfascial ligation.

Forty-seven extremities with recurrent venous ulceration were treated by subfascial ligation of incompetent perforating veins. The limbs were observed for an average of 8.5 years (range, 0.5 to 13 years). The risk for recurrence was 22%, 41%, and 51% at 1, 3, and 5 years, respectively. Patients with bilateral ulceration or prior venous ligation were at the highest risk for recurrence, while those with prior excision of prominent varicose veins had a reduced risk. There has been no loss of limbs or life secondary to this venous problem during the 398 cumulative years of observation.

In situ saphenous vein bypass grafts for limb salvage. A current fad or a viable alternative to reversed vein bypass grafts?

Revascularization of the lower extremity using the in situ saphenous vein bypass graft has resurfaced as a clinical alternative to reversal of the saphenous vein. Early patency rates have been excellent, however, concern has been raised about the durability of the in situ technique. Our total experience with this technique has been reviewed to evaluate its effectiveness on a teaching vascular service. Seventy-six limbs in 71 patients were revascularized using the in situ technique. The distal anastomosis was created at the below-the-knee popliteal level in 26 limbs and at the infrapopliteal level in 50 limbs. Operative assessment of the vein quality showed 42 percent to be phlebitic or less than 4 mm in diameter. Hospital mortality was 0 and late mortality was 8 percent. Cumulative life table analysis showed the graft patency rate to be 89 percent 1 month postoperatively, 82 percent at 1 year, 77 percent at 2 years, and 72 percent up to 4 year postoperatively. Patency was independent of runoff to the pedal arch and the level of the distal anastomosis. Limb salvage at 4 years was 83 percent for distal popliteal grafts and 79 percent for infrapopliteal reconstructions. Our results indicate that the long-term durability of the in situ saphenous vein graft is excellent despite suboptimal veins and poor runoff. When performed properly, it is the preferred technique for arterial reconstruction below the knee joint.

Nickel is a component of hydrogenase in Rhizobium japonicum.

The derepression of H2-oxidizing activity in free-living Rhizobium japonicum does not require the addition of exogenous metal to the derepression media. However, the addition of EDTA (6 microM) inhibited derepression of H2 uptake activity by 80%. The addition of 5 microM nickel to the derepression medium overcame the EDTA inhibition. The addition of 5 microM Cu or Zn also relieved EDTA inhibition, but to a much lesser extent; 5 microM Fe, Co, Mg, or Mn did not. The kinetics of induction and magnitude of H2 uptake activity in the presence of EDTA plus Ni were similar to those of normally derepressed cells. Nickel also relieved EDTA inhibition of methylene blue-dependent Hup activity, suggesting that nickel is involved directly with the H2-activating hydrogenase enzyme. Adding nickel or EDTA to either whole cells or crude extracts after derepression did not affect the hydrogenase activity. Cells were grown in 63Ni and the hydrogenase was subsequently purified by gel electrophoresis. 63Ni comigrated with the H2-dependent methylene blue reducing activity on native polyacrylamide gels and native isoelectric focusing gels. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the nickel-containing hydrogenase band revealed a single polypeptide with a molecular weight of ca. 67,000. We conclude that the hydrogenase enzyme in R. japonicum is a nickel-containing metalloprotein.