Horizontal gene transfer underlies the painful stings of asp caterpillars (Lepidoptera: Megalopygidae).

Larvae of the genus Megalopyge (Lepidoptera: Zygaenoidea: Megalopygidae), known as asp or puss caterpillars, produce defensive venoms that cause severe pain. Here, we present the anatomy, chemistry, and mode of action of the venom systems of caterpillars of two megalopygid species, the Southern flannel moth Megalopyge opercularis and the black-waved flannel moth Megalopyge crispata. We show that megalopygid venom is produced in secretory cells that lie beneath the cuticle and are connected to the venom spines by canals. Megalopygid venoms consist of large aerolysin-like pore-forming toxins, which we have named megalysins, and a small number of peptides. The venom system differs markedly from those of previously studied venomous zygaenoids of the family Limacodidae, suggestive of an independent origin. Megalopygid venom potently activates mammalian sensory neurons via membrane permeabilization and induces sustained spontaneous pain behavior and paw swelling in mice. These bioactivities are ablated by treatment with heat, organic solvents, or proteases, indicating that they are mediated by larger proteins such as the megalysins. We show that the megalysins were recruited as venom toxins in the Megalopygidae following horizontal transfer of genes from bacteria to the ancestors of ditrysian Lepidoptera. Megalopygids have recruited aerolysin-like proteins as venom toxins convergently with centipedes, cnidarians, and fish. This study highlights the role of horizontal gene transfer in venom evolution.

Genome rearrangements induce biofilm formation in Escherichia coli C - an old model organism with a new application in biofilm research.

BACKGROUND: Escherichia coli C forms more robust biofilms than other laboratory strains. Biofilm formation and cell aggregation under a high shear force depend on temperature and salt concentrations. It is the last of five E. coli strains (C, K12, B, W, Crooks) designated as safe for laboratory purposes whose genome has not been sequenced. RESULTS: Here we present the complete genomic sequence of this strain in which we utilized both long-read PacBio-based sequencing and high resolution optical mapping to confirm a large inversion in comparison to the other laboratory strains. Notably, DNA sequence comparison revealed the absence of several genes thought to be involved in biofilm formation, including antigen 43, waaSBOJYZUL for lipopolysaccharide (LPS) synthesis, and cpsB for curli synthesis. The first main difference we identified that likely affects biofilm formation is the presence of an IS3-like insertion sequence in front of the carbon storage regulator csrA gene. This insertion is located 86 bp upstream of the csrA start codon inside the - 35 region of P4 promoter and blocks the transcription from the sigma32 and sigma70 promoters P1-P3 located further upstream. The second is the presence of an IS5/IS1182 in front of the csgD gene. And finally, E. coli C encodes an additional sigma70 subunit driven by the same IS3-like insertion sequence. Promoter analyses using GFP gene fusions provided insights into understanding this regulatory pathway in E. coli. CONCLUSIONS: Biofilms are crucial for bacterial survival, adaptation, and dissemination in natural, industrial, and medical environments. Most laboratory strains of E. coli grown for decades in vitro have evolved and lost their ability to form biofilm, while environmental isolates that can cause infections and diseases are not safe to work with. Here, we show that the historic laboratory strain of E. coli C produces a robust biofilm and can be used as a model organism for multicellular bacterial research. Furthermore, we ascertained the full genomic sequence of this classic strain, which provides for a base level of characterization and makes it useful for many biofilm-based applications.

Drosophila Brainbow: a recombinase-based fluorescence labeling technique to subdivide neural expression patterns.

We developed a multicolor neuron labeling technique in Drosophila melanogaster that combines the power to specifically target different neural populations with the label diversity provided by stochastic color choice. This adaptation of vertebrate Brainbow uses recombination to select one of three epitope-tagged proteins detectable by immunofluorescence. Two copies of this construct yield six bright, separable colors. We used Drosophila Brainbow to study the innervation patterns of multiple antennal lobe projection neuron lineages in the same preparation and to observe the relative trajectories of individual aminergic neurons. Nerve bundles, and even individual neurites hundreds of micrometers long, can be followed with definitive color labeling. We traced motor neurons in the subesophageal ganglion and correlated them to neuromuscular junctions to identify their specific proboscis muscle targets. The ability to independently visualize multiple lineage or neuron projections in the same preparation greatly advances the goal of mapping how neurons connect into circuits.

Surfactant-Impregnated MOF-Coated Fabric for Antimicrobial Applications.

Since the onset of the SARS-CoV-2 pandemic, the world has witnessed over 617 million confirmed cases and more than 6.54 million confirmed deaths, but the actual totals are likely much higher. The virus has mutated at a significantly faster rate than initially projected, and positive cases continue to surge with the emergence of ever more transmissible variants. According to the CDC, and at the time of this manuscript submission, more than 77% of all current US cases are a result of the B.5 (omicron). The continued emergence of highly transmissible variants makes clear the need for more effective methods of mitigating disease spread. Herein, we have developed an antimicrobial fabric capable of destroying a myriad of microbes including betacoronaviruses. We have demonstrated the capability of this highly porous and nontoxic metal organic framework (MOF), γ-CD-MOF-1, to serve as a host for varied-length benzalkonium chlorides (BACs; active ingredient in Lysol). Molecular docking simulations predicted a binding affinity of up to -4.12 kcal·mol-1, which is comparable to that of other reported guest molecules for this MOF. Similar Raman spectra and powder X-ray diffraction patterns between the unloaded and loaded MOFs, accompanied by a decrease in the Brunauer-Emmett-Teller surface area from 616.20 and 155.55 m2 g-1 respectively, corroborate the suggested potential for pore occupation with BAC. The MOF was grown on polypropylene fabric, exposed to a BAC-loading bath, washed to remove excess BAC from the external surface, and evaluated for its microbicidal activity against various bacterial and viral classes. Significant antimicrobial character was observed against Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, bacteriophage, and betacoronavirus. This study shows that a common mask material (polypropylene) can be coated with BAC-loaded γ-CD-MOF-1 while maintaining the guest molecule's antimicrobial effects.

The 44th Medical Brigade in the Great War: Vietnam, 1966-Activation, Deployment, and Initial Operations.

In 1950, Congress changed the name of the Army Medical Department (AMEDD) to the Army Medical Service (AMEDS) as part of the Army Organization Act of 1950. In March 1968, at the urging of Army Surgeon General Leonard D. Heaton, then in his ninth year of service as the Surgeon General, Secretary of the Army Stanley R. Resor petitioned Congress to restore the name of the Army Medical Service to the Army Medical Department, and Congress approved the restoration of the department's name in June 1969.

Bacterial Biofilm Growth on 3D-Printed Materials.

Recent advances in 3D printing have led to a rise in the use of 3D printed materials in prosthetics and external medical devices. These devices, while inexpensive, have not been adequately studied for their ability to resist biofouling and biofilm buildup. Bacterial biofilms are a major cause of biofouling in the medical field and, therefore, hospital-acquired, and medical device infections. These surface-attached bacteria are highly recalcitrant to conventional antimicrobial agents and result in chronic infections. During the COVID-19 pandemic, the U.S. Food and Drug Administration and medical officials have considered 3D printed medical devices as alternatives to conventional devices, due to manufacturing shortages. This abundant use of 3D printed devices in the medical fields warrants studies to assess the ability of different microorganisms to attach and colonize to such surfaces. In this study, we describe methods to determine bacterial biofouling and biofilm formation on 3D printed materials. We explored the biofilm-forming ability of multiple opportunistic pathogens commonly found on the human body including Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus to colonize eight commonly used polylactic acid (PLA) polymers. Biofilm quantification, surface topography, digital optical microscopy, and 3D projections were employed to better understand the bacterial attachment to 3D printed surfaces. We found that biofilm formation depends on surface structure, hydrophobicity, and that there was a wide range of antimicrobial properties among the tested polymers. We compared our tested materials with commercially available antimicrobial PLA polymers.

The accuracy and predictability of micro Doppler radar signature projection algorithm measuring functional movement in NCAA athletes.

BACKGROUND: Development of accessible cost-effective technology to objectively, reliably, and accurately predict musculoskeletal injury risk could aid the effort to prevent chronic pain and disability. Recent work on micro-Doppler radar suggests it merits investigation towards these goals. The micro-Doppler signals that are created can infer differences in gross movements such as walking versus crawling in military settings where direct vision is not possible. Unique micro-Doppler signals may be able to identify more subtle movement patterns which would not be easily seen by the human eye. RESEARCH QUESTION: Can micro Doppler radar predictably and accurately identify subtle differences in movement conditions? METHODS: This is a cross sectional study recruiting NCAA athletes to jump in front of the micro-Doppler radar barefoot, with shoes, and shoes with a heel lift. The micro-Doppler radar signature projection algorithm was developed to determine whether the radar is able to distinguish the three distinct movement patterns. RESULTS: Confusion matrices were used to visualize the performance of the support-vector machine at the 80/20 test/train split correctly classifying barefoot subjects, shoes and heel lift, and shoes correctly at 0° with respect to the radar 90.9 %, 86.7 %, and 89.5 % of the time, respectively. At 90° with respect to the radar, it was successful 94.1 %, 100 %, and 80 % of the time, respectively. CONCLUSION: This study suggests that the micro-Doppler radar signature projection algorithm is highly accurate and able to predict subtle differences in movement that are not readily observed with conventional motion capture systems. Future studies are needed to better understand if micro-Doppler signals can identify pathologic movement patterns or movement that is associated with increased risk of injury.

A search for medications to treat COVID-19 via in silico molecular docking models of the SARS-CoV-2 spike glycoprotein and 3CL protease.

BACKGROUND: The COVID-19 has now been declared a global pandemic by the World Health Organization. There is an emergent need to search for possible medications. METHOD: Utilization of the available sequence information, homology modeling, and in slico docking a number of available medications might prove to be effective in inhibiting the SARS-CoV-2 two main drug targets, the spike glycoprotein, and the 3CL protease. RESULTS: Several compounds were determined from the in silico docking models that might prove to be effective inhibitors for SARS-CoV-2. Several antiviral medications: Zanamivir, Indinavir, Saquinavir, and Remdesivir show potential as and 3CLPRO main proteinase inhibitors and as a treatment for COVID-19. CONCLUSION: Zanamivir, Indinavir, Saquinavir, and Remdesivir are among the exciting hits on the 3CLPRO main proteinase. It is also exciting to uncover that Flavin Adenine Dinucleotide (FAD) Adeflavin, B2 deficiency medicine, and Coenzyme A, a coenzyme, may also be potentially used for the treatment of SARS-CoV-2 infections. The use of these off-label medications may be beneficial in the treatment of the COVID-19.

The Development of a Pipeline for the Identification and Validation of Small-Molecule RelA Inhibitors for Use as Anti-Biofilm Drugs.

Biofilm infections have no approved effective medical treatments and can only be disrupted via physical means. This means that any biofilm infection that is not addressable surgically can never be eliminated and can only be managed as a chronic disease. Therefore, there is an urgent need for the development of new classes of drugs that can target the metabolic mechanisms within biofilms which render them recalcitrant to traditional antibiotics. Persister cells within the biofilm structure may play a large role in the enhanced antibiotic recalcitrance of bacteria biofilms. Biofilm persister cells can be resistant to up to 1000 times the minimal inhibitory concentrations of many antibiotics, as compared to their planktonic envirovars; they are thought to be the prokaryotic equivalent of metazoan stem cells. Their metabolic resistance has been demonstrated to be an active process induced by the stringent response that is triggered by the ribosomally-associated enzyme RelA in response to amino acid starvation. This 84-kD pyrophosphokinase produces the "magic spot" alarmones, collectively called (p)ppGpp. These alarmones act by directly regulating transcription by binding to RNA polymerase. These transcriptional changes lead to a major shift in cellular function to both upregulate oxidative stress-combating enzymes and down regulate major cellular functions associated with growth and replication. These changes in gene expression produce the quiescent persister cells. In this work, we describe a hybrid in silico laboratory pipeline for identifying and validating small-molecule inhibitors of RelA for use in the combinatorial treatment of bacterial biofilms as re-potentiators of classical antibiotics.

[The influence of surgical and restorative dental treatment prior to cardiac valve surgery on the long-term demand of dental treatment: a prospective clinical study]. / Eine klinisch prospektive Untersuchung zum Einfluss der Fokussanierung vor Herzklappenoperation auf den langfristigen zahnärztlichen Behandlungsbedarf.

The study objective was to evaluate the long-term influence of non-radical surgical and restorative dental treatment modalities prior to elective cardiac valve replacement on the subsequent dental treatment demand. A total of 305 patients preceding cardiac valve surgery were screened and the appropriate dental treatment was initiated. After 36 months 80 patients were re-evaluated clinically, of which 60 required dental restorations of 155 teeth, mostly due to periodontal pathology. Independent of the sub-group there was a statistically substantial increase of the treatment demand compared to the time of initial examination. In addition, at the time of final re-evaluation the definite treatment need significantly increased far beyond anticipation, potentially due to inadequate dental procedures during the follow-up interval. Irrespective of any dental treatment or antibiotic application, endocarditis did not occur in any patient. Non-radical dental restoration prior to cardiac valve replacement can only be successful, if a standardized dental follow-up with common monitoring forms is provided. The risk of prosthetic valve infective endocarditis based on a dental focus is probably overestimated.

Need for dental treatment following cardiac valve surgery: a clinical study.

AIM: The aim of the present study was to evaluate the long-term need for dental treatment following non-radical treatment modes prior to cardiac valve surgery. PATIENTS: From 1995 to 2001, a total of 305 patients were screened prior to cardiac surgery. After an average period of 36 months, 80 of these patients could be re-evaluated clinically (26%). Another 117 patients (38%) and their family doctors were contacted by telephone. METHODS: Dental evaluation prior to cardiac valve replacement was performed clinically and radiographically. Tooth extraction was recommended in cases of carious or periodontal destruction, root remnants, partial retention or apical osteolysis despite endodontic treatment with poor prognosis for apicectomy. Periodontal therapy was recommended if attachment loss was less than 1/2 of the root length. No measures were undertaken for endodontically treated teeth without apical osteolysis and impacted teeth. In October 2002, oral health was re-evaluated in 80 patients. Dental treatment carried out in the follow-up period was documented and compared with the current findings. RESULTS: At the time of re-evaluation, 60 of the total of 80 patients required dental treatment in 155 teeth; oral surgery was indicated in 51 of these 60 patients, mostly due to periodontal pathology. During the follow-up period, only 99 of the 142 dental interventions having taken place had been carried out with prophylactic antibiotic treatment. CONCLUSION: From the results of this study it may be concluded that non-radical dental treatment modes prior to cardiac valve replacement can only be successful over the long-term if adequate postoperative dental care is provided. To achieve this aim, common follow-up monitoring forms, similar to those used for care of cancer patients, could facilitate communication.

Long-term outcome after coronary stenting.

The present review assesses the data on long-term outcome after coronary stenting. Histological, angiographical and intravascular imaging data have shown that the insertion of stents constitutes only a transient stimulus to lumen renarrowing, that this process is almost complete at 6 months and that a certain degree of neointima regression is also possible after this time. Clinical data have confirmed the sustained benefit of stenting in the long term. Careful selection of optimal stent designs and application of the recent advances in adjunctive pharmacological therapy are currently effective strategies to improve both short-and long-term results with coronary stenting. However, further efforts are needed and are ongoing to combat restenosis, a process that counters the excellent short-term results of stenting in the long term.

A GAL4-driver line resource for Drosophila neurobiology.

We established a collection of 7,000 transgenic lines of Drosophila melanogaster. Expression of GAL4 in each line is controlled by a different, defined fragment of genomic DNA that serves as a transcriptional enhancer. We used confocal microscopy of dissected nervous systems to determine the expression patterns driven by each fragment in the adult brain and ventral nerve cord. We present image data on 6,650 lines. Using both manual and machine-assisted annotation, we describe the expression patterns in the most useful lines. We illustrate the utility of these data for identifying novel neuronal cell types, revealing brain asymmetry, and describing the nature and extent of neuronal shape stereotypy. The GAL4 lines allow expression of exogenous genes in distinct, small subsets of the adult nervous system. The set of DNA fragments, each driving a documented expression pattern, will facilitate the generation of additional constructs for manipulating neuronal function.

Sex-based analysis of outcome in patients with acute myocardial infarction treated predominantly with percutaneous coronary intervention.

CONTEXT: A higher mortality risk for women with acute myocardial infarction (AMI) is a common finding in studies that compare the postinfarction outcome of women vs men. It is not clear, however, whether sex is an independent predictor of death among patients systematically treated with aggressive reperfusion and medical strategies. OBJECTIVE: To assess the impact of patient's sex on outcome in a consecutive series of patients with AMI treated with a reperfusion strategy largely based on percutaneous coronary interventions. DESIGN, SETTING, AND PATIENTS: Inception cohort of 1937 patients (502 women and 1435 men) who were admitted with a diagnosis of AMI to a tertiary referral institution between January 1995 and December 2000. MAIN OUTCOME MEASURES: Mortality at 1 year after AMI. RESULTS: Compared with men, women were older (70 vs 61 years; P<.001) and had known diabetes or hypertension more often. Both men and women received essentially identical therapy with the majority of patients (86%) receiving reperfusion therapy via percutaneous coronary interventions. There were no significant differences in 1-year Kaplan-Meier death rates with 13.8% (68 cases) among women and 12.9% (184 cases) among men (unadjusted hazard ratio, 1.06; 95% confidence interval, 0.80-1.39; P =.70). After age adjustment, women had a lower risk of death (hazard ratio, 0.65; 95% confidence interval, 0.49-0.87; P =.004). CONCLUSION: Despite their more advanced age and greater prevalence of diabetes or hypertension, women with AMI who were treated with a reperfusion strategy largely based on percutaneous coronary interventions show a similar outcome as men.

Clopidogrel therapy in patients undergoing coronary stenting: value of a high-loading-dose regimen.

The objective of this study was to assess the value of a clopidogrel regimen based on a high loading dose initiated before the stent placement procedure. A consecutive series of 864 patients treated with a high-loading-dose clopidogrel regimen (600 mg given 2-4 hr prior to intervention) was compared with 870 patients treated with conventional ticlopidine therapy. Abciximab was given periprocedurally in 62% of the patients. The composite endpoint of death, myocardial infarction, or urgent revascularization was reached by 39 (4.5%) clopidogrel patients and 59 (6.8%) ticlopidine patients. Clopidogrel therapy was associated with a 35% reduction of the risk for early adverse events (odds ratio 0.65; 95% confidence interval, 0.43-0.98). Thus, a high-loading-dose clopidogrel regimen in patients undergoing coronary artery stenting was safe and led to a more favorable clinical outcome than conventional therapy with ticlopidine regardless of concomitant treatment with abciximab.

Characterization and biological activity of a Brazilian isolate of Bacillus sphaericus (Neide) highly toxic to mosquito larvae

Primary powders of Bacillus spharicus strain S2 isolated from soil samples in Brazil, and strain 2362 were produced in a 14 liter fermentor. Growth patterns and sporulation observed in three trials with strains S2 and 2362 in the fermentor were similar. Second-instar larvae of Culex quinquefasciatus, Anopheles albimanus, Anophles quadrimaculatus and Aedes aegypti exposed for 48 hr to strain S2 responded with LC50 values of 0.25, 5.95, 12.28 and 140.0 ppb of lyophilized primary powder, respectively. Under the same conditions, strain 2362 resulted in LC50 values of 0.39, 7.16, 16.93 and 307.0 ppb of lyophilized primary powder, respectively, in those mosquito larvae. Statistical analysis of the bioassay data did not show significant differences among LC50 values observed in B. sphaericus strains S2 and 2362, at the 0.05 level. Toxins of strains S2 and 2362 were extracted at pH 12 with NaOH. Electrophoresis of the extraxts in plyacrylamide gel under denaturing conditions reveled the 51 and 42 kDa toxins in both S2 and 2362 B. Sphaericus strains. The presence of the 42 Da peptide in the extracts was confirmed by Western blot and Elisa, with anti-42 kDa IgG previously prepared from strain 2362.