Honey bee promoter sequences for targeted gene expression.

The honey bee, Apis mellifera, displays a rich behavioural repertoire, social organization and caste differentiation, and has an interesting mode of sex determination, but we still know little about its underlying genetic programs. We lack stable transgenic tools in honey bees that would allow genetic control of gene activity in stable transgenic lines. As an initial step towards a transgenic method, we identified promoter sequences in the honey bee that can drive constitutive, tissue-specific and cold shock-induced gene expression. We identified the promoter sequences of Am-actin5c, elp2l, Am-hsp83 and Am-hsp70 and showed that, except for the elp2l sequence, the identified sequences were able to drive reporter gene expression in Sf21 cells. We further demonstrated through electroporation experiments that the putative neuron-specific elp2l promoter sequence can direct gene expression in the honey bee brain. The identification of these promoter sequences is an important initial step in studying the function of genes with transgenic experiments in the honey bee, an organism with a rich set of interesting phenotypes.

[Medical honey in the treatment of wound-healing disorders in the head and neck area]. / Erste Erfahrungen mit medizinischem Honig in der Wundbehandlung im Kopf-Hals-Bereich.

BACKGROUND: Already in ancient times honey was used as a drug and for the treatment of wounds. In recent years the different effects of honey on wound-healing processes have been reexamined. Based on this, the antibacterial and fungicidal qualities of honey could be confirmed. PATIENTS AND METHODS: Between January 2009 and July 2011 medical honey was used on 36 patients suffering from different wound-healing disorders in the head and neck area after unsuccessful conventional treatment. The healing process was registered by microbiological investigations, measurements of the wound edges and adequate photo documentation. RESULTS: Medical honey can be used without problems or detectable side effects on problematic wounds of the head and neck area. Constant treatment leads to fast wound lavation, granulation, reduction of putrid smells and a decrease in inflammation. CONCLUSIONS: In the treatment of poorly healing and infected wounds within the head and neck area, medical honey can be used successfully without problems as an effective alternative to conventional treatment options.

Learning channels. Cellular physiology of odor processing neurons within the honeybee brain.

To understand the cellular mechanisms of olfactory learning in the honeybee brain we study the physiology of identified neurons within the olfactory pathway. Here, we review data on the voltage-sensitive and ligand-gated ionic currents of mushroom body Kenyon cells and antennal lobe neurons in vitro and in situ. Both cell types generate action potentials in vitro, but have different voltage-sensitive K+ currents. They express nicotinic acetylcholine receptors and ionotropic GABA receptors, representing the major transmitter systems in the insect olfactory system. Our data are interpreted with respect to learning-dependent plasticity in the honeybee brain.

Morphology of feedback neurons in the mushroom body of the honeybee, Apis mellifera.

The anatomy of gamma-aminobutyric acid (GABA)-immunoreactive, recurrent feedback neurons in the mushroom body (MB) of the honeybee, Apis mellifera, was investigated by using intraneuropilar injections of cobalt ions and light microscopic techniques. Each MB contains approximately 110 GABA-immunoreactive neurons, and approximately 50% of them are feedback neurons, i.e., they connect the MB output regions--the alpha-lobe, beta-lobe, and pedunculus--with its input regions--the calyces. Their somata are located in the lateral protocerebral lobe, and their primary neurites project medially and bifurcate near the alpha-lobe. In the alpha-lobe feedback neurons form narrow banded, horizontal arborizations in the dorsal and median alpha-lobe; each cell innervates a certain alpha-lobe layer. The neurons form additional branches in the pedunculus and the beta-lobe. All calycal subcompartments--the lip, collar, and basal ring--are innervated by feedback neurons. However, individual feedback neurons innervate exclusively a certain subcompartment in both the median and lateral calyx. Due to the arrangement of intrinsic Kenyon cells, each calycal subcompartment is connected to its specific, corresponding layer in the alpha-lobe. Feedback neurons interconnect the alpha-lobe and the calyces in either a corresponding or a noncorresponding fashion. With respect to their branching pattern in the alpha-lobe, the basal ring and the collar neuropil receive input from feedback neurons innervating the corresponding dorsal and median alpha-lobe layers. By contrast, the lip region, which receives olfactory antennal input, is innervated by feedback neurons with arborizations in a noncorresponding dorsal alpha-lobe layer.

Nicotinic acetylcholine currents of cultured Kkenyon cells from the mushroom bodies of the honey bee Aapis mellifera.

1. Acetylcholine-induced currents of mushroom body Kenyon cells from the honey bee Apis mellifera were studied using the whole-cell configuration of the patch clamp technique. Pressure application of 1 mM acetylcholine (ACh) induced inward currents with amplitudes between -5 and -500 pA. 2. The cholinergic agonists ACh and carbamylcholine had almost equal potencies of current activation at concentrations between 0.01 and 1 mM; nicotine was less potent. The muscarinic agonist oxotremorine did not elicit any currents. 3. Approximately 80 % of the ACh-induced current was irreversibly blocked by 1 microM alpha-bungarotoxin. Atropine (1 mM) did not block the ACh-induced current. 4. Upon prolonged ACh application the current desensitized with a time course that could be approximated by the sum of two exponentials (tau1 = 276 +/- 45 ms (mean +/- s.e.m. ) for the fast component and tau2 = 2.4 +/- 0.7 s for the slow component). 5. Noise analyses of whole-cell currents yielded elementary conductances of 19.5 +/- 2.4 pS for ACh and 23.7 +/- 5.0 pS for nicotine. The channel lifetimes, calculated from the frequency spectra, were tauo = 1.8 ms for ACh and tauo = 2.5 ms for nicotine. 6. Raising the external calcium concentration from 5 to 50 mM shifted the reversal potential of the ACh-induced current from +4. 6 +/- 0.9 to +37.3 +/- 1.3 mV. The calcium-to-sodium permeability ratio (PCa : PNa) was 6.4. 7. In high external calcium solution (50 mM) the ACh-induced current rectified in an outward direction at positive membrane potentials. 8. We conclude that Kenyon cells express nicotinic ACh receptors with functional profiles reminiscent of the vertebrate neuronal nicotinic ACh receptor subtype.

Ecdysteroid control of ionic current development in Manduca sexta motoneurons.

The steroid hormone 20-hydroxyecdysone (20-HE) regulates several processes during insect metamorphosis. We studied the effects of 20-HE on the development of voltage-sensitive ionic currents of thoracic leg motoneurons of Manduca sexta. The larval leg motoneurons persist throughout metamorphosis but undergo substantial morphological reorganization, which is under the control of 20-HE and accompanied by changes in Ca2+ and K+ current densities. To determine whether 20-HE controls the changes in Ca2+ and K+ current levels during postembryonic development, identified thoracic leg motoneurons isolated from late larval and early pupal stages were taken into primary cell culture. Whole-cell Ca2+ and K+ currents were measured after 1-4 days of steroid hormone incubation. In the presence of 20-HE, peak Ca2+ currents of pupal leg motoneurons increased from day 1 to day 4 in vitro. Thus, at culture day 4 the pupal Ca2+ current levels were larger in 20-HE-treated than in untreated cells. By contrast, 20-HE did not affect the Ca2+ current amplitudes of larval leg motoneurons. Whole-cell K+ currents, measured at 4 days in pupal motoneurons, consisted of a fast-activating transient current and a sustained, slowly inactivating current. 20-HE did not affect the amplitude of the transient or sustained currents after 4 days in vitro. Thus, a direct steroid hormone effect may control the proper maturation of voltage-sensitive Ca2+ currents in leg motoneurons.

Zebrafish mutations affecting retinotectal axon pathfinding.

We have isolated mutants in the zebrafish Danio rerio that have defects in axonal connectivity between the retina and tectum. 5-day-old fish larvae were screened by labeling retinal ganglion cells with DiI and DiO and observing their axonal projections to and on the tectum. 82 mutations, representing 13 complementation groups and 6 single allele loci, were found that have defects in retinal ganglion cell axon pathfinding to the tectum. These pathfinding genes fall into five classes, based on the location of pathfinding errors between eye and tectum. In Class I mutant larvae (belladonna, detour, you-too, iguana, umleitung, blowout) axons grow directly to the ipsilateral tectal lobe after leaving the eye. Class II mutant larvae (chameleon, bashful) have ipsilaterally projecting axons and, in addition, pathfinding mistakes are seen within the eye. In Class III mutant larvae (esrom, tilsit, tofu) fewer axons than normal cross the midline, but some axons do reach the contralateral tectal lobe. Class IV mutant larvae (boxer, dackel, pinscher) have defects in axon sorting after the midline and retinal axons occasionally make further pathfinding errors upon reaching the contralateral tectal lobe. Finally, Class V mutant larvae (bashful, grumpy, sleepy, cyclops, astray) have anterior-posterior axon trajectory defects at or after the midline. The analysis of these mutants supports several conclusions about the mechanisms of retinal axon pathfinding from eye to tectum. A series of sequential cues seems to guide retinal axons to the contralateral tectal lobe. Pre-existing axon tracts seem not to be necessary to guide axons across the midline. The midline itself seems to play a central role in guiding retinal axons. Axons in nearby regions of the brain seem to use different cues to cross the ventral midline. Mutant effects are not all-or-none, as misrouted axons may reach their target, and if they do, they project normally on the tectum. The retinotectal pathfinding mutants reveal important choice points encountered by neuronal growth cones as they navigate between eye and tectum.

Mutations disrupting the ordering and topographic mapping of axons in the retinotectal projection of the zebrafish, Danio rerio.

Retinal ganglion cells connect to their target organ, the rectum, in a highly ordered fashion. We performed a large-scale screen for mutations affecting the retinotectal projection of the zebrafish, which resulted in the identification of 114 mutations. 44 of these mutations disturb either the order of RGC axons in the optic nerve and tract, the establishment of a topographic map on the tectum, or the formation of proper termination fields. Mutations in three genes, boxer, dackel and pinscher, disrupt the sorting of axons in the optic tract but do not affect mapping on the tectum. In these mutants, axons from the dorsal retina grow along both the ventral and the dorsal branch of the optic tract. Mutations in two genes, nevermind and who-cares, affect the dorsoventral patterning of the projection. In embryos homozygous for either of these mutations, axons from dorsal retinal ganglion cells terminate ventrally and dorsally in the tectum. In nevermind, the retinotopic order of axons along the optic nerve and tract is changed in a characteristic way as well, while it appears to be unaffected in who-cares. Two mutations in two complementation groups, gnarled and macho, affect the anteroposterior patterning of the projection. In these mutants, nasodorsal axons branch and terminate too soon in the anterior tectum. In 27 mutants belonging to six complementation groups, retinal axons do not form normal termination fields. Some implications for models concerning the formation of topographic projections are discussed.

The ocelli of arctiid moths: ultrastructure of the retina during light and dark adaptation.

The ultrastructure of the dorsal ocelli of two arctiid moths (Arctia caja (A. caja) and Creatonotos transiens (C. transiens) was investigated. The two ocelli are positioned laterally on the vertex of the head posterior to the antennae, close to the dorsal margin of the compound eyes. The biconvex corneal lens is located at the apex of a cone-shaped cuticular elevation, which encapsulates the retina. The corneagenous cell layer and the cup-like retina with about 100-130 receptor cells in A. caja (70-90 receptor cells in C. transiens) are adjoined proximally. The retina is completely enclosed by the perineurium and thus separated from the corneagenous cells and the surrounding hemolymph. Irregularly shaped rhabdomeres, consisting of densely packed microvilli, are present in the distal region of the receptor cells. Up to three cells may form a rhabdom. Thus a loose network of photoreceptive structures over the whole retina results. A unique feature of these arctiid ocelli are photoreceptor vacuoles containing microvilli. The function of these organelles is unknown. The rhabdomeric arrangement within the light and dark adapted retina differs considerably. The ultrastructure of the rhabdomeres indicates an intense membrane turnover. However, changes in adaptation state are not accompanied by dramatic changes in the photoreceptive area of an ocellus.

Hernia uteri (correction of: uterus) inguinale associated with unilateral renal agenesis.

We report a patient with a rudimentary uterine horn, fallopian tube and ovary in an inguinal hernia. Associated with this abnormality the patient had ipsilateral renal agenesis and a unicornuate uterus.

A role for the mesoderm in endodermal migration and morphogenesis in Drosophila.

The endodermal midgut arises from two primordia, the anterior midgut (AMG) primordium and the posterior midgut (PMG) primordium, which are separated by almost the entire length of the Drosophila embryo. To form the midgut, these two parts have to extend towards each other and to fuse laterally on both sides of the yolk. Shortly before and during that movement, AMG and PMG are arranged as mesenchymal cell masses, but later the midgut cells form an epithelium. We show that these two aspects of midgut development, migration of AMG and PMG and transition to an epithelium, depend on the mesoderm. The extension of the midgut primordia is achieved by cell migration along the visceral mesoderm which forms a continuous layer of cells within the germ band. In mutant embryos lacking the entire mesoderm or failing to differentiate the visceral mesoderm, AMG and PMG are formed but do not migrate properly. In addition, they fail to form an epithelium and instead either remain as compact cell masses anterior and posterior to the yolk (in twist and snail mutant embryos) or only occasionally wrap around the yolk before embryogenesis is completed (in tinman-deficient embryos). We conclude that the visceral mesoderm serves as a substratum for the migrating endodermal cells and that the contact between visceral mesoderm and endoderm is required for the latter to become an epithelium.

Radiology of starch granulomatous peritonitis.

Starch granulomatous peritonitis (SGP) is a rare complication of abdominal surgery that results, in a susceptible individual, from contamination of the peritoneal cavity with starch particles from the surgeon's gloves. It is important to make the diagnosis of SGP without recourse to re-operation as the optimal treatment is non-surgical. A case of SGP that was confirmed only after re-operation and resection of the involved tissues is reported. The first description of an abdominal computed tomography scan in this condition is presented, the significance of which was only recognized postoperatively.

Should the 'normal' appendix be removed at operation for appendicitis?

A prospective study was conducted over a 6-month period to assess the ability of the surgeon to determine the presence or absence of acute inflammation of the appendix at operation for acute appendicitis. Over the study period 175 appendices were removed of which 131 (75%) were acutely inflamed histologically. The surgeon's assessment as to the presence of acute inflammation was correct in 120 out of 132 cases, a positive predictive value of 91%. When the surgeon assessed the appendix as being non-inflamed he was wrong in 11 of 43 cases, a false reassurance rate of 26%. The increasing use of laparoscopy in the assessment and treatment of patients with the clinical features of appendicitis should not alter the practice of removing the appendix when it appears macroscopically normal. To do so may result in a number of patients requiring a second operation to remove the appendix in the early postoperative period.

Asynchronous ovarian torsion--the case for prophylactic oophoropexy.

The case of an 11 year old girl who suffered torsion of the left ovary 8 months following right oophorectomy for a similar event is presented. The left ovary was saved by immediate surgical intervention. The value of prophylactic oophoropexy in unilateral ovarian torsion is discussed.

Induction of 2'-5' oligoadenylate synthetase during interferon treatment of chronic myelogenous leukemia.

Activity of the interferon-induced enzyme 2'-5' oligoadenylate synthetase (2-5 OAS) was measured in peripheral blood mononuclear cells (PBMCs) and serum of patients with chronic phase Ph'-positive chronic myelogenous leukemia (CML) treated with interferon-alpha (IFN-alpha) (4 x 10(6) IU/m2) alone or in combination with 50 micrograms IFN-gamma. At the beginning of IFN therapy, marked elevation of 2-5 OAS titers was detected in PBMCs (pretreatment 0.03-1.62, median 0.2; during treatment 0.8-13.14, median 4.31; 22 patients studied) and in serum (pretreatment 21-156 pmol/dl, median 62; during treatment 532-1740 pmol/dl, median 800; eight patients studied). However, 2-5 OAS titers were not related to clinical outcome or IFN therapy and also during IFN resistance elevated 2-5 OAS activity in PBMCs (median 3.45; range 1.05-13.14; 11 patients studied) were detected. These data argue against direct involvement of the 2-5 OAS system in the therapeutic effect of IFN in CML. However, 2-5 OAS titers in PBMCs or serum appear to be a reliable control of biologically active IFN therapy.