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Biomedical subjects

James E Baker

Publications and source records attributed to James E Baker.

13 recordsLinked to original sources

RNAi-induced silencing of embryonic tryptophan oxygenase in the Pyralid moth, Plodia interpunctella.

Gene silencing through the introduction of double-stranded RNA (RNA interference, RNAi) provides a powerful tool for the elucidation of gene function in many systems, including those where genomics and proteomics are incomplete. The use of RNAi technology for gene silencing in Lepidoptera has lacked significant attention compared to other systems. To demonstrate that RNAi can be utilized in the lepidopteran, Plodia interpunctella, we cloned a cDNA for tryptophan oxygenase, and showed that silencing of tryptophan oxygenase through RNAi during embryonic development resulted in loss of eye-color pigmentation. The complete amino acid sequence of Plodia tryptophan oxygenase can be accessed through NCBI Protein Database.

Amino Acid Sequence↗

Phenoloxidase in larvae of Plodia interpunctella (Lepidoptera: Pyralidae): molecular cloning of the proenzyme cDNA and enzyme activity in larvae paralyzed and parasitized by Habrobracon hebetor (Hymenoptera: Braconidae).

Phenoloxidase (PO) is a major component of the insect immune system. The enzyme is involved in encapsulation and melanization processes as well as wound healing and cuticle sclerotization. PO is present as an inactive proenzyme, prophenoloxidase (PPO), which is activated via a protease cascade. In this study, we have cloned a full-length PPO1 cDNA and a partial PPO2 cDNA from the Indianmeal moth, Plodia interpunctella (Hubner) (Lepidoptera: Pyralidae) and documented changes in PO activity in larvae paralyzed and parasitized by the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae). The cDNA for PPO1 is 2,748 bp and encodes a protein of 681 amino acids with a calculated molecular weight of 78,328 and pI of 6.41 containing a conserved proteolytic cleavage site found in other PPOs. P. interpunctella PPO1 ranges from 71-78% identical to other known lepidopteran PPO-1 sequences. Percent identity decreases as comparisons are made to PPO-1 of more divergent species in the orders Diptera (Aa-48; As-49; and Sb-60%) and Coleoptera (Tm-58; Hd-50%). Paralyzation of host larvae of P. interpunctella by the idiobiont H. hebetor results in an increase in phenoloxidase activity in host hemolymph, a process that may protect the host from microbial infection during self-provisioning by this wasp. Subsequent parasitization by H. hebetor larvae causes a decrease in hemolymph PO activity, which suggests that the larval parasitoid may be secreting an immunosuppressant into the host larva during feeding.

Amino Acid Sequence↗

Insect fragments in flour: relationship to lesser grain borer (Coleoptera: Bostrichidae) infestation level in wheat and rapid detection using near-infrared spectroscopy.

We determined that the number of insect fragments, quantified using the standard flotation method, in flour milled from wheat infested with larvae, pupae, or preemergent adults of the lesser grain borer, Rhyzopertha dominica (F.), was proportional to infestation level. Wheat infested with a single preemergent adult contributed 28 and 10x as many fragments as wheat infested with a single larva or pupa, respectively. Using regression models that were developed from these data, we predicted that the maximum infestation level that would result in flour with fragment counts below the Food and Drug Administration defect action level (75 fragments/50 g of flour) was 0.95 and 1.5% (380-640 infested kernels/kg of wheat) for pupae and larvae, but it decreased to 0.05% (20 infested kernels/kg) when the grain was infested with preemergent adults. We also reexamined the accuracy and sensitivity of near-infrared spectroscopy (NIRS) for detecting insect fragments in flour by testing three different NIR spectrometers. NIRS-predicted numbers of insect fragments were correlated with the actual number of fragments. NIRS is less precise than the standard flotation method, but it is rapid, nondestructive, does not require extensive sample preparation, and could easily be automated for a more sophisticated sampling protocol for flour based on prescreening samples with NIRS followed up by use of the standard flotation method when necessary.

Animals↗

Innate immunity in a pyralid moth: functional evaluation of domains from a beta-1,3-glucan recognition protein.

Invertebrates, like vertebrates, utilize pattern recognition proteins for detection of microbes and subsequent activation of innate immune responses. We report structural and functional properties of two domains from a beta-1,3-glucan recognition protein present in the hemolymph of a pyralid moth, Plodia interpunctella. A recombinant protein corresponding to the first 181 amino-terminal residues bound to beta-1,3-glucan, lipopolysaccharide, and lipoteichoic acid, polysaccharides found on cell surfaces of microorganisms, and also activated the prophenoloxidase-activating system, an immune response pathway in insects. The amino-terminal domain consists primarily of an alpha-helical secondary structure with a minor beta-structure. This domain was thermally stable and resisted proteolytic degradation. The 290 residue carboxyl-terminal domain, which is similar in sequence to glucanases, had less affinity for the polysaccharides, did not activate the prophenoloxidase cascade, had a more complicated CD spectrum, and was heat-labile and susceptible to proteinase digestion. The carboxyl-terminal domain bound to laminarin, a beta-1,3-glucan with beta-1,6 branches, but not to curdlan, a beta-1,3-glucan that lacks branching. These results indicate that the two domains of Plodia beta-1,3-glucan recognition protein, separated by a putative linker region, bind microbial polysaccharides with differing specificities and that the amino-terminal domain, which is unique to this class of pattern recognition receptors from invertebrates, is responsible for stimulating prophenoloxidase activation.

Animals↗

Stage-specific surface chemicals of Plodia interpunctella: 2-acyl-1,3-cyclohexanediones from larval mandibular glands serve as cuticular lipids.

Cuticular lipid compositions of all life stages of the stored product moth Plodia interpunctella have been determined. Eggs and adults of P. interpunctella have cuticular lipids consisting solely of hydrocarbons. The composition of eggs and adult females is qualitatively nearly identical with ca. 86 hydrocarbons (11 n-alkanes, 39 monomethyl alkanes, 19 dimethyl alkanes, 11 trimethyl alkanes and 6 monoenes) except females lack the 2-methyl alkanes found in eggs. Adult males have a hydrocarbon composition qualitatively nearly identical to females with the exception that they lack the monoenes. Larval and pupal cuticular lipids are dominated by a mixture of ca. 20 previously described 2-acyl-1,3-cyclohexanediones, with only minute amounts of n-alkanes on the larvae and pupae. The 2-acyl-1,3-cyclohexanediones are continuously secreted onto their silk webbing and food particles by the paired mandibular glands found in all larvae. Extracts from dissected mandibular glands have a qualitatively identical composition to larval cuticular extracts. The pupal stage (which does not have mandibular glands) is enclosed in a silk cocoon also coated with 2-acyl-1,3-cyclohexanediones laid down while the wandering stage larvae spin the cocoon. The 2-acyl-1,3-cyclohexanediones have physical properties which closely mimic those of cuticular hydrocarbons, including melting point and boiling point range and hydrophobicity. This is the first report of an insect with a life stage that does not use conventional cuticular lipids for conservation of water.

Animals↗

Profound hypoxemia resulting from shunting across an inadvertent atrial septal tear after left ventricular assist device placement.

UNLABELLED: Defects within the interatrial septum (IAS) can be a source of significant right-to-left shunting and hypoxemia, particularly after placement of a left ventricular assist device (LVAD). We report a case of LVAD placement in which an unrecognized IAS tear occurred intraoperatively, leading to profound arterial desaturation. Transesophageal echocardiography (TEE) was instrumental in making the diagnosis. Certain intraoperative events increased the pressure gradient between the right and left atria, aggravating hypoxemia. We recommend that patients undergoing LVAD placement be screened intraoperatively with TEE for unrecognized IAS defects. Re-examination of the IAS should occur on weaning from cardiopulmonary bypass. IMPLICATIONS: A traumatic atrial septal defect after atrial cannulation caused a right-to-left intracardiac shunt on initiation of left ventricular assist device support that was further aggravated by chest closure and pleural suction, culminating in severe hypoxemia.

Adult↗

Chronological age-grading of three species of stored-product beetles by using near-infrared spectroscopy.

The accuracy of near-infrared spectroscopy (NIRS) for predicting the chronological age of adults of the rice weevil, Sitophilus oryzae (L.); the lesser grain borer, Rhyzopertha dominica (F.); and the red flour beetle, Tribolium castaneum (Herbst), three pests of stored grain, was examined. NIRS-predicted age correlated well with actual age of these three species. Age predictions in S. oryzae by using the NIRS method are not dependent upon adult sex or temperatures to which adult weevils are exposed. Results indicated that water content decreased with increasing age in rice weevil adults, and excluding wavelengths at which water absorbs NIR radiation reduced the accuracy of correct classification. Additionally, removing cuticular lipids from insects resulted in a significant decrease in classification accuracy of weevils, indicating that these compounds may be partly responsible for the ability of NIRS to differentiate young from old beetles. NIRS is a nondestructive technique that can be used to age-grade large numbers of adult stored-product beetles, information that could help to increase the accuracy of population models for these pest species.

Animals↗

Cuticular hydrocarbons and wax esters of the ectoparasitoid Habrobracon hebetor: Ontogenetic, reproductive, and nutritional effects.

Hydrocarbon and wax ester components of cuticular lipids of the braconid parasitoid Habrobracon hebetor Say reared at 25 degrees C on larvae of a pyralid moth have been identified by GC-MS and analyzed with respect to adult age, mating status, and diet. The hydrocarbons range in carbon number from C(21) to C(45) and consist of a homologous series of n-alkanes, 11-, 13-, and 15-methyl alkanes, 13,17-dimethyl alkanes, and Z-5, Z-7, and Z-9-alkenes. The wax esters found in the cuticular lipid fraction are a series of homologous compounds with the acid portion being short chain, unbranched, even carbon number acids from C(8) to C(20) (predominately C(8) to C(16)). The alcohol portions of the esters are secondary alcohols with carbon number from C(22) to C(25) (predominately C(23) and C(25)) with the hydroxyl function located at C(6), C(7), C(8), and C(9). Gender, age, and nutritional states were significant factors for variation in several of the individual esters, but mating status did not affect wax ester composition. Ontogenetic examinations indicated that prepupal, and early pupal cuticular lipids contain only hydrocarbons. Low levels of wax esters are detectable in late stage pupae, and somewhat greater quantities of wax esters are present on newly eclosed adults. When pharate adults emerge from the cocoon, however, their cuticular lipids consist of approximately equal amounts of hydrocarbons and wax esters, and 6d post emergence from the cocoon, wax esters are the predominant lipid component.

Age Factors↗

Novel diterpenoids and hydrocarbons in the Dufour gland of the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae).

Chemical constituents contained in the Dufour gland of the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae) were characterized. Three terpenes, beta-springene, a homo-beta-springene, and a homo-geranyllinalool constitute approximately 37% of the gland components, with the remaining 63% all being hydrocarbons. The hydrocarbons consist of a homologous series of n-alkanes (n-C21 to n-C31), a trace amount of 3-methyl C23, a homologous series of internally methyl-branched alkanes (11-methyl C23 to 13-methyl C35), one dimethylalkane (13,17-dimethyl C33), a homologous series of monoenes (C(25:1) to C(37:1)) with the double bonds located at Delta9, Delta13 and Delta15 for alkenes of carbon number 25 to 31 and at Delta13 and Delta15 for carbon numbers 33 to 37 and three homologous dienes in very low amounts with carbon numbers of 31, 32, and 33. The terpenoid and hydrocarbon composition of the Dufour gland was similar in virgin and mated females. However, in contrast to the hydrocarbons, the amount of beta-springene and homo-geranyllinalool increased significantly with time after adult emergence from the cocoon. Although many hydrocarbons in the Dufour gland are the same as those on the cuticle of this species [Howard and Baker, Arch. Insect Biochem. Physiol. 53:1-18 (2003)], substantial differences also occur. Of particular note is the chain length of alkenes and location of the double bonds: cuticular alkenes have a chain length of C23 to C29 and double bond locations at Delta5, Delta7, and Delta9, whereas the Dufour gland alkenes contains a greater range of carbon numbers and have no Delta5 or Delta7 alkenes. The Dufour gland contains only one of the long-chain dimethylalkanes found on the cuticle. Also, no terpenoids are found on the cuticle, and the Dufour gland contains none of the secondary wax esters that are major components on the cuticle. GC-MS analysis of lipids carried in the hemolymph of H. hebetor indicated that all hydrocarbons found on both the cuticle and in the Dufour gland are present, as are some of the wax esters. However, none of the terpenoids were detected in the hemolymph. This suggests that the hydrocarbons are synthesized in other tissues or cells, probably by oenocytes, and differentially partitioned between the cuticle and the Dufour gland. The terpenoids are most likely synthesized within the Dufour gland. Analysis of surface lipids from eggs laid within 18 h indicated that no diterpenoids were present. Rather, the lipids present on the eggs were n-alkanes, monomethylalkanes, alkenes, and secondary alcohol wax esters. This composition did not reflect that of the Dufour gland, hence eggs are not being coated with Dufour gland components during oviposition.

Animals↗

Morphology and chemistry of Dufour glands in four ectoparasitoids: Cephalonomia tarsalis, C. waterstoni (Hymenoptera: Bethylidae), Anisopteromalus calandrae, and Pteromalus cerealellae (Hymenoptera: Pteromalidae).

The venom apparatus of four hymenopterous parasitoids, including two bethylids, C. tarsalis (Ashmead) and C. waterstoni (Gahan), and two pteromalids, A. calandrae (Howard) and P. cerealellae (Ashmead), were removed and the associated Dufour glands characterized with respect to their external morphology and chemistry. Dufour glands in all four species have a characteristic translucent appearance that apparently results from their lipid content. The stalked Dufour glands of C. tarsalis and C. waterstoni are pear-shaped and have overall lengths of approximately 0.2 and 0.15 mm, respectively. The thin venom glands are bifurcate and insert through a fine duct into the transparent ovoid- to pear-shaped venom reservoir in these bethylids. In A. calandrae and P. cerealellae the Dufour glands are elongated, tubular structures of ca. 0.35 and 0.8 mm in length, respectively, that constrict to a short stalk that empties into the common oviduct. The venom glands in these pteromalids are simple elongated structures that insert into the sac-like venom reservoir through a fine duct. The chemistry of the volatile contents of the Dufour gland in these four species differs considerably. C. tarsalis Dufour glands contain the same hydrocarbon components as found on the cuticle of this species (Ann. Entomol. Soc. Am. 91:101-112 (1998)), and no other chemicals. The Dufour glands of C. waterstoni also contain only hydrocarbons, most of which are the same as the cuticular hydrocarbons (Ann. Entomol. Soc. Am. 85:317-325 (1992)), but in addition the Dufour gland contains ca. 3% of a mixture of 2,17- and 2,19-dimethyl C(23). A. calandrae Dufour gland chemistry is somewhat more complex than that of either of the two bethylids, but like the bethylids, only hydrocarbons are present. The carbon number range is from C(30) to C(39) and consists of a mixture of n-alkanes (C(30)-C(38)); 3-, 5-, 7-, 9-, 11-, 12-, 13-, 14-, 15- and 17-methyl alkanes; 3,7- and 3,11-dimethyl alkanes; 5,9- and 5,17-dimethyl alkanes; 7,11-, 9,13-, 13,17-, 14,18- and 15,19-dimethyl alkanes; 3,7,11- and 3, 9,15-trimethyl alkanes; and 3,7,11,15-tetramethyl alkanes. The cuticular hydrocarbons of this species have not been previously reported, but they are the same as the Dufour gland hydrocarbons. The Dufour glands of P. cerealellae contain both hydrocarbons and two long-chain aldehydes. Most of the hydrocarbons are identical to those found on the cuticle of this species (Ann. Entomol. Soc. Am. 94:152-158 (2001)), but in addition, 5,9-dimethyl C(27), 5,13-, 5,17- and 5,19-dimethyl C(35), 12- and 14-methyl C(36), 12,16- and 13,17-dimethyl C(36), 13-methyl C(37) and 13,17-dimethyl C(37) are present. The two aldehydes detected in glands from P. cerealellae are n-tetracosanal (C(23)CHO) and n-hexacosanal (C(25)CHO).

Animals↗

Molar pregnancy.

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Adult↗

Chronological age-grading of house flies by using near-infrared spectroscopy.

The sensitivity and accuracy of near-infrared spectroscopy (NIRS) was compared with that of the pteridine fluorescence technique for estimating the chronological age of house flies, Musca domestica (L.). Although results with both techniques were significantly correlated with fly age, confidence limits on predicted ages generally were smaller with NIRS. Young flies could be readily differentiated from old flies by using NIRS. Age predictions using the pteridine method are dependent upon size, sex, and temperature at which adult flies are exposed. In contrast, those factors do not need to be determined for age-grading using NIRS. Classification accuracy using the NIRS method was similar for whole flies, fresh heads, dried heads, and ethanol-preserved heads. The NIRS method was also suitable for predicting age of stable flies, Stomoxys calcitrans (L.), and face flies, Musca autumnalis De Geer. NIRS has several advantages over the measurement of pteridine levels for age-grading field-collected flies, including speed and portability of instrumentation, and not needing to determine sex, size, and temperatures to which adult flies were exposed.

Animals↗