Tickcidal effect of monoclonal antibodies against hemocytes, Om21, in an adult female tick, Ornithodoros moubata (Acari: Argasidae)

In the present study, monoclonal antibodies (mAbs) against adult Ornithodoros moubata hemocytes were established. Afterward, artificial feeding was performed to assess the tickcidal effect of fetal bovine serum meal containing each mAb. As a result, Om21 showed the strongest tickcidal effect on adult female O. moubata. The reactivity of various tick cells and organs, including the hemocyte, midgut, trachea, ovary, fat body, and muscle, to Om21 was then examined by an indirect immunofluorescent antibody test and by immunoelectron microscopy. Om21 reacted with not only hemocytes but also with fat body cells, epidermis, cuticle of the trachea, connective tissue of the muscle, and the basement membrane of the midgut, trachea, fat body, oocyte, and epidermis. These results suggest that Om21 passing through the midgut epithelium induced a tickcidal effect on hemocytes or various organs. However, the target of Om21 could not be identified in the present study. The antihemocyte mAb produced in this study, Om21, may be useful for the immunological control of ticks.     

The distribution of the neural fat body sheath and the accessibility of the extraneural space in the stick insect, Carausius morosus

The fluid compartment delimited at the surface of the central nervous system by the closely applied fat body sheath has been shown to be accessible to molecules of horseradish peroxidase and Indian ink particles. Quantitative analysis indicates that leakage via the intercellular spaces between fat body cells would preclude the possibility of a significant involvement of the fat body cells in sodium regulation of the extraneural fluid. This possibility is further precluded by the absence of a fat body sheath from peripheral nerves.     

结缔组织铺片脂肪组织油红染色在组织学实验教学中的应用

目的在传统结缔组织铺片基础上开展脂肪组织油红染色方法在医学本科生组织学实验教学中的应用。方法学生先进行疏松结缔组织铺片,并施行脂肪组织油红o-甲苯胺兰-伊红三重染色,然后镜下观察。结果油红o染色把结缔组织中的脂肪细胞内脂滴保存下来并染上红色。脂肪组织中央的细胞脂滴均匀红染,充满胞浆,周边的脂肪细胞胞浆中油红染色很少,细胞呈空泡状,显示出脂肪细胞亚群存在。甲苯胺兰染色使得疏松结缔组织中肥大细胞染成紫红色,胞核染色浅,细胞数量多、成群分布。伊红可使得结缔组织内除脂肪细胞、肥大细胞意外的其他细胞的胞浆和胶原纤维染成淡红色。结论传统的组织学平铺片技术基础上引入脂肪油红o-甲苯胺兰-伊红三重染色,可增强学生动手能力,并能很好地了解输送结缔组织中细胞的不同表型和分布,丰富组织学内容,把教学、科研连接一起,达到提高实验教学质量的目的。     

Pathology of Spiroplasma floricola in Galleria mellonella larvae

Spiroplasma floricola strain 23-6, originally isolated from tulip tree flowers, was injected into larvae of the greater wax moth. Histopathology and cytopathology of disease larvae were studied by histochemical, fluorescent antibody, and electron microscopical methods. The gut was empty, polysaccharides in fat and muscle tissue were reduced, the fat body was broken down, and phospholipids were depleted in larvae 4 days after injection. Fluorescein conjugated S. floricola antibody was adsorbed onto hemocytes, sarcolemma, gut epithelial membrane, and the cortex of the ventral ganglia. By electron microscopy, spiroplasmas were found in hemocoel, hemocytes, pericardial cells, connective tissues, basement membranes, epidermal cells of the cuticle, the neural lamella, and the peripheral glial cells of the ventral nerve cord, and on midgut and epidermal membranes. It is postulated that the cytopathological effects induced in the body of the insect released nutritional elements that allow extensive reproduction of S. floricola.     

Intracellular freezing, viability, and composition of fat body cells from freeze-intolerant larvae of Sarcophaga crassipalpis.

Although it is often assumed that survival of freezing requires that ice formation must be restricted to extracellular compartments, fat body cells from freeze-tolerant larvae of the gall fly, Eurosta solidaginis (Diptera, Tephritidae) survive intracellular freezing. Furthermore, these cells are highly susceptible to inoculative freezing by external ice, undergo extensive lipid coalescence upon thawing, and survive freezing better when glycerol is added to the suspension medium. To determine whether these traits are required for intracellular freeze tolerance or whether they are incidental and possessed by fat body cells in general, we investigated the capacity of fat body cells from nondiapause-destined and diapause-destined (i.e., cold-hardy) larvae of the freeze-intolerant flesh fly Sarcophaga crassipalpis (Diptera, Sarcophagidae) to survive intracellular freezing. Fat body cells from both types of larvae were highly susceptible to inoculative freezing; all cells froze between -3.7 to -6.2 degrees C. The highest rates for survival of intracellular freezing occurred at -5 degrees C. The addition of glycerol to the media markedly increased survival rates. Upon thawing, the fat body cells showed little or no lipid coalescence. Fat body cells from E. solidaginis had a water content of only 35% compared to cells from S. crassipalpis larvae that had 52-55%; cells with less water may be less likely to be damaged by mechanical forces during intracellular freezing.     

Fine structural alterations with age in the fat body of the adult male housefly,Musca domestica

Summary The fat body of the adult housefly is composed of two types of cells, the lipid-and glycogen-rich fat body cells and the oenocytes. A comparison of the fine structure of the abdominal fat body in 4-day old and 31–35 day old male houseflies indicated an increase in lipid and a decrease in glycogen content in the fat body cells of old flies. Oenocytes of old flies exhibit deteriorative alterations with an accumulation of secondary lysosomes. Both fat body cells and oenocytes in senile flies are ingested by hemocytes.Supported by a grant from the National Science Foundation.     

Biosynthesis of major plasma proteins in the primary culture of fat body cells from the silkworm, Bombyx mori

Plasma proteins termed "SP1" and "30K proteins" are synthesized by the fat body cells of the silkworm, Bombyx mori, in a sex- and stage-specific manner during larval development. We successfully established a primary culture of the fat body cells in order to investigate the regulatory mechanisms of plasma protein gene expression. The primary cultures of fat body cells contained at least two cell types: small oval cells, and large spherical cells. The cells adhered to and migrated on the cultured dish after plating. By the 7th day of cultivation, the cells clustered to form fat body-like structures, which were maintained for at least 3 months. Plasma proteins were actively synthesized in the primary cultures of the fat body cells isolated from the final instar larvae only when the cells tightly adhered to and clustered on the cultured dish. Immunocytochemical analysis revealed that only 10-15% of the clustered cells synthesized plasma proteins in our culture system, indicating that the primary culture comprises heterogeneous cells that are morphologically and functionally distinct. The patterns of SP1 syntheses in primary cultures faithfully reproduced their sex-dependency in vivo.     

Inhibition of the formation of protein storage granules by glutaurine in the larval fat body cells of Mamestra brassicae (Insecta, Lepidoptera)

Correlative changes in the protein contents of haemolymph and fat body and the accumulation of protein storage granules in the fat body cells of Mamestra brassicae were investigated during the last larval stage in normally developing larvae and following administration of glutaurine (1 X 10(-4) mg/g body weight). The protein content of the haemolymph of untreated larvae increased up to the 4th day of the stage, declined during days 5 and 6, and increased again before pupation. In the glutaurine-treated larvae the amount of proteins in the haemolymph was as high as in the controls during the first four days but continued to rise up to the end of the stage. The protein content of the fat body started to increase from the 3rd day and heavy accumulation of protein storage granules in the cells of fat body was observed on the 5th and following days. The protein content of the fat body of glutaurine-treated larvae remained at a low level and the protein storage granules were absent in the cells. The inhibition of the selective uptake of haemolymphatic storage proteins by fat body following glutaurine treatment is suggested.     

Secretion and Uptake of 14C Proteins by Fat Body of Calpodes ethlius Stoll. (Lepidoptera, Hesperiidae)

Insect fat body from larvae of Calpodes ethlius synthesised proteins in vitro , some of which were secreted into the medium. ¹⁴C leucine was incorporated into fat-body proteins and the released proteins were characterised by electrophoresis and immunodiffusion. The released proteins included the three main proteins normally found in larval hemolymph and fat body. Larval fat body in the main synthetic phase of development did not sequester these ¹⁴C-labelled proteins when incubated with them in vitro or when they were injected into mid-instar larvae. In contrast, these proteins were sequestered by fat-body cells undergoing pupation.
The conclusion is that larval fat-body cells in the main synthetic phase selectively exclude proteins synthesised in vitro by fat body of the same age. During pupal development the fat body absorbs proteins it secreted at an earlier age.     

Macromastia: how much of it is fat?

A total of 25 patients who underwent bilateral breast reduction were included in this study. Each patient's age, weight, height, and amount of breast tissue removed from each breast were recorded. The body mass index was calculated for each patient. On the day of the operation, tissue samples (two each) were taken from the central, lateral, and preaxillary areas of the breast. One of the samples was weighed, placed in a closed glass container, and heated for 10 minutes in a microwave oven at full power. The liquid fat was separated from the solid residue, and the percentage of fat was calculated. The other sample from each area was examined grossly, and representative sections, corresponding to the distribution of fat and connective tissue, were submitted for evaluation. In these samples, the percentage of fat, gland, and connective tissue was estimated using low-magnification light microscopy. In this group of patients (who had an average age of 34 years and who were significantly overweight as determined by a mean body mass index of 28), it was found (using the microwave method) that there was a mean fat percentage of 61 percent in the central breast area, 74 percent in the lateral breast area, and 73 percent in the preaxillary area. Upon microscopic examination, the pathologist reported that fat accounted for 64 percent of the central breast area, 92 percent of the lateral breast area, and 94 percent of the preaxillary area. On average, the central breast area in macromastia patients had only seven percent gland and 29 percent connective tissue. The lateral and preaxillary areas of the breast had one to three percent gland and five percent connective tissue. The two methods had a significant (p < 0.05) positive correlation in the central breast area, but in the lateral and preaxillary regions, the correlation was poor. In the microscopic examination, there was a tendency to overestimate the amount of fat. Both methods of evaluation used in the study concur that the enlarged breast of macromastia consists primarily of fat and that the glandular element is rather small.     

Regional and functional differentiation in the fat body of pharaoh's ant queens, Monomorium pharaonis (L.)

The insect fat body is generally described as a uniform tissue with multiple functions, but we have found evidence of cell differentiation in the Monomorium fat body. We show that the fat body of a mature egg-laying pharaoh's ant queen is a result of a preceding remodeling of cell material comprising at least 11 different fat cell types, located at specific positions in the head, alitrunk (thorax) and gaster (abdomen). The cell types are classified based on their position, histochemistry, ultrastructure, and immunoreactivity for vitellogenin/vitellin. Some of these cells are primordial cells present at emergence, others invade the histolysing flight muscle tissue, and still others disappear during the maturation process. Only one type, the subepidermal fat cell of the gaster, is active in vitellogenin synthesis and is the only cell type in close association with oenocytes. Although only this type produces vitellogenin, our material indicates that most fat cell types are essential to support egg production. In some queens vitellogenin was found to form crystals in ventral vitellogenin-producing fat cells. This indicates an imbalance between vitellogenin production in the fat cells and uptake in the oocytes, which is probably related to a cyclic regulation of egg production.     

Properties,synthesis, and accumulation of storage proteins in Pieris rapae L.

Two kinds of storage proteins (SP-1, SP-2) were confirmed in hemolymph and fat body of Pieris rapae during metamorphosis. Both proteins were present in high concentrations in the hemolymph during the last larval instar. Hemolymph concentrations of SP-1 and SP-2 dropped after pupation as the proteins were being deposited in fat bodies. SP-2 is present in a larger amount than SP-1. Detailed studies on storage proteins determined their properties, mode of synthesis, and accumulation in the fat body. SP-1 has a molecular weight of 500,000 and consists of one type of subunit (M_r 77,000), while SP-2 has a molecular weight of 460,000 and is composed of two types of subunits (M_r 80,000 and 69,000). The pl values of SP-1 and SP-2 were determined to be 6.97 and 7.06, respectively. Fat body cells from 1-day-old fifth instar larvae synthesized storage proteins in large amounts, whereas those from late prepupae exhibited high protein sequestration. Proteins taken up in fat body accumulated in dense granules during the pupal stage but sharply decreased at the adult stage. Morphological changes in the fat body tissues were observed during the larval-pupal transformation; the nuclei of fat body cells became irregularly shaped, and the boundaries between cells seemed to be obscure. Synthesis, storage, or degradation of storage proteins in fat body during development is closely associated with morphological changes in the tissues.     

Effects of carcass maturity on meat quality characteristics of beef semitendinosus muscle for chinese native yellow steers

This work was designed to study the effects of carcass maturity on meat quality characteristics and intramuscular connective tissue of beef semitendinosus muscle from Chinese native Yellow steers. Chemical determinations, histological and mechanical measurements were performed on the raw and cooked meat at 4 days post mortem. In raw meat, intramuscular fat, collagen solubility, mechanical strength and transition temperature of intramuscular connective tissue increased (P < 0.05) with carcass maturity before body maturation, whilst moisture, total collagen, fibre diameter decreased after body maturation. Warner-Bratzlar shear force (WBSF) of cooked meat increased with maturity before body maturation due to the muscle atrophy, and thus the decline of moisture content and the increase of cooking losses. After body maturation, the increase of WBSF was neutralised by the increase of intramuscular fat, the decrease of total collagen and the elongation of sarcomere length.     

Induction of hemolin gene expression by bacterial cell wall components in eri-silkworm, Samia cynthia ricini

A cDNA clone encoding hemolin was isolated from fat body of immunized Samia cynthia ricini larvae based on subtractive suppression hybridization method. The cDNA encodes 413 amino acid residue open reading frame with an 18 residue predicted signal peptide. The expression of the gene was strongly induced in fat body and midgut by an injection of bacterial cells or peptidoglycans, but very weakly by lipopolysaccharide. The mRNA expression in the fat body was detected as early as 3 h post-injection, and reached the peak level at 12 h.     

Body size and composition of Crabeater seals (Lobodon carcinophagus), with observations on tissue and organ size in Ross seals (Ommatophoca rossi)

Linear body measurements and body weights of 17 Crabeater seals and four Ross seals were recorded, and the relationships of weight to linear dimensions were calculated. There were no significant differences between sexes of these relationships in Crabeater seals. All Ross seals were males.
The major body components (blood, fat, skin, muscle, bone, connective tissue and viscera) of seven Crabeater seals were weighed after dissection.
Blood, fat and skin of two Ross seals were weighed. Weights of 22 visceral organs of the same animals, and linear bone dimensions of eight Crabeater seals and skull measurements of five Ross seals were recorded.
There was no significant difference between sexes or ages in body composition of Crabeater seals. Relatively, Crabeater and Ross seals had less blood (9–10% body weight) than Elephant seals, and less fat (21–22% body weight) than most other marine mammals. The low body fat content may have been attributable to season and physiological status of the animals when dissected. The percentages of body weight represented by the other major components of Crabeater seals were: skin 8%, muscle 44%, bone 10%, connective tissue 0.7% and total viscera 8%. These figures, and the relative sizes of individual organs, were discussed in relation to their possible function in Crabeater and Ross seals.     

Larval fat body cells die during the early pupal stage in the frame of metamorphosis remodelation in Bombyx mori

In holometabolus insects, morphology of the larval fat body is remodeled during metamorphosis. In higher Diptera, remodeling of the fat body is achieved by cell death of larval fat body cells and differentiation of the adult fat body from primordial cells. However, little is known about remodeling of the fat body at pupal metamorphosis in Lepidoptera. In this study, we found that cell death of the larval fat body in Bombyx mori occurs at shortly after pupation. About 30% of the fat body cells underwent cell death on days 1 and 2 after pupation. The cell death involved genomic DNA fragmentation, a characteristic of apoptosis. Surgical manipulation and in vitro culture of fat body cells revealed that 20-hydroxyecdysone and juvenile hormone had no effect on either initiation or progression of cell death. During cell death, a large increase in activity of caspase-3, a key enzyme of cell death, was observed. Western blot analysis of the active form of caspase-3-like protein revealed that the length of caspase-3 of B. mori was much larger than that of caspase-3 in other species. The results suggest that larval fat body cells of B. mori are removed through cell death, which is mediated by a caspase probably categorized in a novel family.     

Juvenile hormone and the ultrastructural properties of the fat body of the adult Colorado beetle,Leptinotarsa decemlineata say

Summary In the fat body of ovipositing female Colorado beetles, two types of lobes occur. The first type, the internal fat body, is highly specialised for protein synthesis. A lobe of the second type, the peripheral fat body, contains two types of cells, oenocytes and glycogen cells. Ovariectomy, performed at adult moult results in hypertrophy of the glycogen cells of the peripheral fat body. The lobes are characterized by the storage of lipid bodies and glycogen and by numerous mitochondria. Short-day conditions ab ovo, which induce diapause in adults, also result in hypertrophy of glycogen cells of the peripheral fat body. Furthermore, only few mitochondria occur but many proteinaceous bodies may be observed, which conditions are in contrast to the observed effects of castration. The fat body of allatectomized long-day females, has the same structure as that of short day beetles. Consequently a lack of juvenile hormone induces the proteinaceous bodies.Dr. A. De Loof gratefully acknowledges a scholarship as Aspirant of the National Foundation of Scientific Research in Belgium. We wish to thank Prof. Dr. h. C. J. de Wilde for his suggestions and helpfull criticism. We also thank Mr. W. Bohijn for his help in operating the EM and Mr. G. Maes for photography.     

Regional differentiation of fat bodies in larvae of the indianmeal moth,Plodia interpunctella

Larvae of the Indianmeal moth, Plodia interpunctella, contain two morphologically distinct fat bodies. Tan-colored, highly tracheated fat body located posteriorly in the abdomen was the predominant fat body tissue during the early larval instars. White, sheet fat body located more anteriorly became the predominant type during the fifth (last) larval instar and eventually occupied most of the space of the hemocoel. Ultrastructural morphology of tan fat body showed the tissue to be composed of cells containing numerous, large, spherical mitochondria, with only few lipid, glycogen, or protein storage structures. In contrast, white fat body was composed of cells that in later larval stages had organelles typical of storage functions. Both fat bodies produced storage proteins during the late fifth instar, whereas only white fat body accumulated the storage proteins. Tan fat body dispersed and apparently autolyzed in pharate pupae, whereas the white fat body metamorphosed and persisted into the adult stage. These observations indicate that fat body of the Indianmeal moth is functionally and morphologically differentiated along the anterior-posterior axis into two regional subgroups of cells.     

Fat body cells of Amblyomma cajennense partially engorged females (Acari: Ixodidae) and their role on vitellogenesis process

During the process of Arthropoda reproduction, the synthesis and uptake of proteins, carbohydrates, and lipids by oocytes is termed vitellogenesis. These compounds that will make up the yolk may be in ticks endogenously synthesized by the oocytes and/or exogenously produced by the fat body and pedicel cells. This study examined the fat body of Amblyomma cajennense ticks at the cytochemical ultrastructural level to demonstrate the presence of lipids, proteins and carbohydrates in trophocytes. The lipids were detected in higher quantity than proteins and carbohydrates in the fat body cells, suggesting that the role of the fat body in tick is stored lipids and carbohydrates to convert them in energy, or still they could be used with cell structural purpose. The electrophoresis technique applied at A. cajennense fat body demonstrated specifically the molecular mass of proteins: about 98 kDa. By the other hands, the fat body is not the organ responsible for the synthesis of the yolk protein, role probably performed by the pedicel cells.