Allergic fungal rhinosinusitis: detection of fungal DNA in sinus aspirate using polymerase chain reaction.

OBJECTIVE: This study investigated allergic fungal rhinosinusitis cases, and aimed to compare the detection of fungi in sinus aspirate by culture and by polymerase chain reaction assay, and to relate the presence of fungi in the nasal sinuses to the type of fungal allergen causing disease. METHODS: Sixty-eight cases of allergic fungal rhinosinusitis underwent fungal culture and polymerase chain reaction assay for universal fungal, aspergillus and bipolaris DNA. Aspergillus-specific immunoglobulin E levels were measured in sinus aspirate, and total serum immunoglobulin E levels were calculated. A control group of 10 cases was included in the study. RESULTS: Of the 68 allergic fungal rhinosinusitis cases, only 42 (61.7 per cent) had positive fungal cultures; of the 10 controls, only three (30 per cent) had positive cultures. Species from the dematiaceous family were most commonly grown, being isolated in 30 cases (71.4 per cent). Bipolaris was the most commonly isolated species (18 cases) followed by curvularia (11 cases) and alternaria (one case). Polymerase chain reaction assay detected fungal DNA in all the allergic fungal rhinosinusitis cases and also in four controls (40 per cent). Ten patients (of 68; 14.7 per cent) were positive for Aspergillus fumigatus specific immunoglobulin E. The mean concentration of this immunoglobulin was 11.32 +/- 4.12 IU/ml in patients and 0 IU/ml in controls, a statistically significant difference. CONCLUSION: Detection of fungal DNA in nasal aspirate by polymerase chain reaction was superior to fungal cultures as a method of detecting fungal growth. In allergic fungal rhinosinusitis, fungal growth is not always accompanied by an allergic reaction.

Clinical and microbiological efficacy of MDT in the treatment of diabetic foot ulcers.

OBJECTIVE: To assess the clinical and microbiological efficacy of maggot debridement therapy (MDT) in the management of diabetic foot ulcers unresponsive to conventional treatment and surgical intervention. METHOD: Consecutive diabetic patients with foot wounds presenting at the vascular surgery unit and the diabetic foot unit of Alexandria Main University Hospital were selected for MDT. Lucilia sericata medicinal maggots were applied to the ulcers for three days per week. Changes in the percentage of necrotic tissue and ulcer surface area were recorded each week over the 12-week follow-up period. Semiquantitative swab technique was used to determine the bacterial load before and after MDT. RESULTS: The sample comprised 10 patients with 13 diabetic foot ulcers. The mean baseline ulcer surface area was 23.5cm2 (range 1.3-63.1), and the mean percentage of necrotic tissue was 74.9% (range 29.9-100). Complete debridement was achieved in all ulcers in a mean of 1.9 weeks (range 1-4). Five ulcers (38.5%) were completely debrided with one three-day MDT cycle. The mean reduction in ulcer size was significant at 90.2%, and this occurred in a mean of 8.1 weeks (range 2-12). The mean weekly reduction in ulcer size was 16.1% (range 8.3-50). Full wound healing occurred in 11 ulcers (84.6%) within a mean of 7.3 weeks (range 2-10). The bacterial load of all ulcers reduced sharply after the first MDT cycle to below the 10(5) threshold, which facilitates healing. CONCLUSION: The results highlight the potential benefits of MDT in diabetic wound care in developing countries. MDT was proved to be a rapid, simple and efficient method of treating these ulcers.

Ultrastructural observations on the elusive subtegumental cells of the viviparous gill monogenean, Macrogyrodactylus clarii.

The ultrastructure of the tegument, including putative subtegumental cells, of the monogenean gill parasite Macrogyrodactylus clarii is described for the first time. The lateral cells are secretory in nature and can be classified into four types: T1 producing elongated, electron-dense, bodies (t1); T2 manufacturing electron-lucent vesicles (t2); T3 producing thin, dump-bell shaped, electron-dense bodies (t3); and T4 containing lyzosome-like inclusions. Morphologically, the T2 cells resemble those described as presumptive vitelline cells in other gyrodactylids whereas the T1, T3 and T4 cells are reported for the first time in gyrodactylids. The close similarity between the secretory inclusions of the outer syncytial layer of the tegument and those found in the T1, T2 and T3 cells suggests that these cells represent the enigmatic tegumental cell bodies rather than vitelline cells. The possible functions of the distinctive features of the tegument of M. clarii are discussed.

Neuromusculature of Macrogyrodactylus congolensis, a monogenean skin parasite of the Nile catfish Clarias gariepinus.

Phalloidin fluorescence technique, enzyme cytochemistry and immunocytochemistry, in conjunction with confocal scanning laser microscopy, were used to describe the neuromusculature of the monogenean skin parasite Macrogyrodactylus congolensis from the Nile catfish Clarias gariepinus. The body wall muscles are composed of an outer layer of compactly arranged circular fibres, an intermediate layer of paired longitudinal fibres and an inner layer of well-spaced bands of diagonal fibres arranged in two crossed directions. The central nervous system consists of paired cerebral ganglia from which three pairs of longitudinal ventral, lateral and dorsal nerve cords arise. The nerve cords are connected at intervals by many transverse connectives. Both central and peripheral nervous systems are bilaterally symmetrical and better developed ventrally than laterally and dorsally. Structural and functional correlates of the neuromusculature of the pharynx, haptor and reproductive tracts were examined. Results implicate acetylcholine, FMRFamide-related peptides and serotonin in sensory and motor function. The results were compared with those of Macrogyrodactylus clarii, a gill parasite of the same host fish C. gariepinus.

Neuromusculature of Macrogyrodactylus clarii, a monogenean gill parasite of the Nile catfish Clarias gariepinus in Egypt.

Phalloidin fluorescence technique, enzyme cytochemistry and immunocytochemistry in conjunction with confocal scanning laser microscopy have been used for the first time to describe the nervous and muscle systems of the viviparous monogenean gill parasite, Macrogyrodactylus clarii. The gross spatial arrangement of muscle and associated cholinergic, peptidergic and aminergic innervations has been examined. The central nervous system (CNS) consists of paired cerebral ganglia from which emanate three pairs of longitudinal ventral, lateral and dorsal nerve cords, connected at intervals by transverse connectives. The CNS is better developed ventrally than dorsally or laterally, and has the strongest reactivity for all neuroactive substances examined. Structural and functional correlates of the neuromusculature of the pharynx, haptor and reproductive tracts have been examined. Results implicate acetylcholine, FMRFamide-related peptides (FaRPs) and serotonin in sensory and motor function in this monogenean, although confirmatory physiological data are obviously required.

Identification of 100 KDa protein in sera of mice-treated with Cu(II) complex with superoxide dismutase-mimetic activity.

The protein profile of sera isolated from mice pre-treated with Cu(II) complex of Girard T with superoxide dismutase (SOD)-mimetic activity was analyzed using SDS-PAGE. This complex was intraperitoneally administered (10 mg/Kg body weight) to Swiss albino mice. The resolved polypeptides showed a new sharp band at 100 KDa against which a polyclonal antibody was raised in rabbit. Sera of rabbit anti-100 KDa protein was used as a powerful probe for the detection of 100 KDa protein isolated from sera of treated mice. Western blot assays revealed a strong reactive polypeptide band at 100 KDa in sera of the mice, but no cross reaction was observed with sera of normal mice. The identification of purified polypeptide was confirmed by different characterization experiments.

Two bellies of the coracobrachialis muscle associated with a third head of the biceps brachii muscle.

Reports that describe the abnormalities and complexities of the anatomy of the arm are important with regard to surgical approaches. This case study reports a combined abnormal form of the coracobrachialis and biceps brachii muscles of the left arm of an adult male cadaver that was detected during the educational gross anatomy dissections of embalmed cadavers. The coracobrachialis muscle demonstrated two bellies which formed shortly inferior to its origin from the coracoid process of the scapula. One belly inserted into the middle of the antero-medial surface of the humerus, whereas the other belly inserted into the medial head of the triceps brachii muscle. The musculocutaneous nerve passed between the two bellies, giving a separate branch to each. We suggest that the two bellies of the coracobrachialis muscle may represent the incompletely fused short heads of the ancestral muscle. The biceps brachii muscle showed a third head, which originated mainly from the antero-medial surface of the humerus and partially from an aponeurosis belonging to the medial head of the triceps brachii muscle. These observations were confined to the left upper limb and were not accompanied by any other abnormality.

Chaetotaxy of the monogeneans Macrogyrodactylus clarii and m. congolensis from the gills and skin of the catfish clarias gariepinus in Egypt, with a note on argentophilic elements in the nervous system.

A comparison has been made between the chaetotaxy of the gyrodactylid monogeneans Macrogyrodactylus clarii Gussev, 1961 and M. congolensis (Prudhoe, 1957) Yamaguti, 1963 from the gills and skin, respectively, of the catfish Clarias gariepinus (Burchell) from the river Nile in Egypt. Bilaterally arranged argentophilic structures on the surface of these parasites are presumed to be sensilla and are more abundant in M. clarii than in M. congolensis especially on the ventral surface (124 vs. 66). In both species these sensilla are concentrated on the head lobes and in the pharyngeal region, but there are features of the sensilla patterns that can be used to distinguish the two species. Comparison is made with sensilla patterns of other gyrodactylids. A system of cells and dendritic processes, most probably part of the nervous system, also has an affinity for silver in the two species. There are no previous records of extensive argentophilic elements in the nervous systems of monogeneans.

A study on the morphology of the coracobrachialis muscle and its relationship with the musculocutaneous nerve.

Thirty-six arms from embalmed adult cadavers were utilised for this investigation. Coracobrachialis muscle was carefully examined to record variations in its attachments, morphology and its relationship with the musculocutaneous nerve. The results of the present work identified the presence of two heads of origin for the coracobrachialis muscle, which are situated superficial (anterior) and deep (posterior) to the musculocutaneous nerve. The superficial head arises mainly from most of the medial border of the tendon of the short head of the biceps brachii muscle. The deep head arises from the apex of the coracoid process of the scapula and the adjoining lateral border of the tendon of the short head of the biceps brachii muscle. The musculocutaneous nerve provides a separate branch for each head. In cases where the coracobrachialis muscle was not pierced by the musculocutaneous nerve, the muscle was formed of one head that has an origin analogous to that of the superficial head. One case showed a splitting of the deep head into two bellies shortly after its origin, where the muscle appeared as being formed of three heads. Variations in the insertion were present as an additional aponeurotic insertion above the usual insertion and an aponeurotic extension to the deep fascia on the medial aspect of the arm. Variations in the musculocutaneous nerve were in the form of lower origin from the lateral root of median nerve and a nerve with a short course after which it united with the median nerve. This investigation supplied evidence of the double heads of origin for the coracobrachialis muscle. The detected variability in insertion and association of the muscle with the musculocutaneous nerve further supports the idea that the coracobrachialis muscle is a complex muscle.

Ultrastructural immunogold study on the various cell types of cultured pancreatic islets of adult rats.

Whereas several reports describing the ultrastructure of the intact pancreatic islets have been recorded, published experience with the ultrastructural integrity of the cultured pancreatic islets is limited. The present study was, therefore, undertaken to provide an ultrastructure identification of the different cells in the cultured islets of the adult rat pancreas, after marking their secretory granules with gold particles. Pancreatic islets were isolated from adult male Wistar rats by the intraductal perfusion of collagenase technique. The islets were cultured in RPMI-1640 medium for 3 days and processed for preparation of ultrathin sections. The sections were stained with the indirect immunogold technique for insulin, glucagon, somatostatin, and pancreatic polypeptide. Ultrastructural examination of the cultured islets clearly identified the presence of B, A, D and PP-cells, as indicated by the numerous gold particles concentrated predominantly over the secretory granules. The secretory granules of the various cell types of the cultured islets demonstrated several similarities as well as differences from the recorded results of the corresponding secretory granules of the intact islets. The differences probably reflect a deviation in the underlying mechanisms of synthesis, maturation and secretion of the different secretory products of the cells in the cultured islets as they adapt to the in vitro environment.

Expression of interferon-gamma, interleukin-4 and interleukin-5 mRNA in the nasal mucosal membrane of rats with allergic rhinitis.

The production of immunoglobulin E (IgE) antibody is largely dependent on the ratio between interleukin-4 (IL-4) (a T helper 2 (Th2)-type cytokine) and interferon-gamma (IFN-gamma) (a T helper 1 (Th1)-type cytokine). Interleukin-5 (IL-5) (also a Th2-type cytokine) is an important eosinophil differentiation factor and also co-stimulates B-cell growth and differentiation. The present study was designed to evaluate and compare the expression of IFN-gamma, IL-4 and IL-5 mRNA in the nasal mucosal membrane of sensitized Brown-Norway (BN) rats. Fourteen BN rats were divided into two groups: non-sensitized (control) and sensitized. The sensitized group was injected with ovalbumin (OA) intraperitoneally on three consecutive days. Twenty-one days later, rats were exposed to 1% OA aerosol. Twenty-four hours after exposure to aerosol, nasal mucosa was extracted from both groups and reverse transcriptase-polymerase chain reaction (RT-PCR) was performed. The densities of the bands of IL-4, IL-5 and IFN-gamma mRNA were expressed as percentages against beta-actin mRNA. Our results showed that the mean values for IL-4 and IL-5 mRNA were increased significantly in sensitized rats compared with control rats. In contrast, the mean value for IFN-gamma mRNA was significantly lower in sensitized rats compared with those of the control group. Our data therefore suggest that sensitization of rat nasal mucous membranes results in the predominant expression of Th2-type cytokines.

Effect of allergen-specific immunotherapy on interleukin-4, interleukin-5 and interferon-gamma mRNA expression in the nasal mucosa of rats with allergic rhinitis.

To elucidate the mechanism of immunotherapy, we tested the effect of ovalbumin and ovalbumin-pullulan conjugate immunotherapy on the expression of interleukin (IL)-4, IL-5 and interferon-gamma (IFN-gamma) mRNA in the nasal mucosa of sensitized rats. Forty-five rats were injected with ovalbumin intraperitoneally on three consecutive days and later were exposed to ovalbumin aerosol. The rats were injected intradermally, on six consecutive days, with saline, ovalbumin or ovalbumin-pullulan conjugate. Later, nasal mucosa was obtained and reverse transcription-polymerase chain reaction (RT-PCR) was performed. Nasal responses and specific immunoglobulin E (IgE) were measured. Although the immunotherapy significantly decreased nasal airway resistance, dye leakage and histamine content in nasal irrigation after allergen challenge, no significant difference was found in IL-4 and IL-5 mRNA expression or in specific IgE level among the three groups. We conclude that in this allergic model, the improvement of nasal responses after immunotherapy was the result of a mechanism other than decrease of T-helper 2 (Th2) cytokines.

Antitumor activities of vanadium(IV), manganese(IV), iron(III), cobalt(II) and copper(II) complexes of 2-methylaminopyridine.

The effect of Cu(II), Mn(IV), Fe(III), V(IV) and Co(II) complexes of 2-methylaminopyridine (L) having superoxide dismutase (SOD)-like activities on Ehrlich ascites carcinoma (EAC) cells was studied. Each of these complexes was intraperitoneally administered (10 mg/kg body weight for 9 days) to Swiss albino mice implanted intraperitoneally with 1 x 10(6) EAC cells. Six days after the last treatment the EAC cells were harvested using a heparinized syringe. The volume of EAC cells and EAC cell viability as well as changes in the levels of tumor cell enzyme activities of SOD, catalase, glutathione peroxidase (GSH-Px), glutathione reductase (GSH-R) and glucose-6-phosphate dehydrogenase (G6PD) were tested to examine the antitumor effects of these complexes. Both tumor volume and tumor cell viability were significantly lowered in complex-treated mice. After tumor transplantation and treatment with the complexes, the activities of GSH-Px and GSH-R were significantly lowered while SOD and G6PD activities were increased in EAC cells compared to their levels in EAC cells harvested from saline-treated mice.

Protective action of some Cu(II) complexes against photohemolysis induced by m. chloroperbenzoic acid.

m. Chloroperbenzoic acid (CPBA) was used as a photosensitizer in the photolysis of human red blood cells. Treatment of the cells with 25, 50 100, or 200 microM CPBA produced 3.5%, 85.9%, 252.6% or 350.9% photohemolysis, respectively, compared with untreated control cells. Some copper (II) complexes [CuL2] with SOD-like activity containing 2-methyl amino pyridine and various anions (Cl-, Br-, NO3-, ClO4-, or SO4--) were tested for their protective efficiency toward the cell damage photoinduced by CPBA. Treatment of the cells with maximum dose (200 microM) Cu(II), bromide, chloride, nitrate, sulfate, or perchlorate complexes with 2-methyl amino pyridine produced 85.87%, 59.28%, 49.29%, 33.47%, or 23.80% protection, respectively, against the cells irradiated without addition of Cu(II) complexes. A mechanism involving oxygen radicals and sensitizer photodegradation products was suggested to interpret the results.

Superoxide dismutase activities in serum and white blood cells of patients with some malignancies.

Superoxide dismutase (SOD) activity has been estimated in serum and leukocytes isolated from adults and children with acute lymphoblastic leukemia (ALL), non-Hodgkin lymphoma (NHL), Hodgkin's disease (HD) and neuroblastoma. Superoxide dismutase levels in leukocytes isolated from adult patients with HD, ALL and NHL were significantly higher than the corresponding control values. Serum SOD activity of adult patients with ALL and NHL was significantly decreased compared to the normal value, while its activity was not significantly changed in patients with HD. Marked deficiencies of leukocyte SOD levels in children with ALL, NHL, HD and neuroblastoma were found. There were no apparent differences in serum SOD activity in children suffering from ALL, NHL, HD and neuroblastoma. It is concluded that the assay of leukocyte SOD can be used to differentiate between HD and NHL in both adults and children.

Some urinary enzymes in bilharziasis.

Urinary alkaline phosphatase (ALP), acid phosphatase (ACP), aryl sulphatase (Ar. sulph.), beta-glucuronidase (beta-gluc.) and galactosidase were assayed in a group of Bilharzia haematobium patients and another group of healthy subjects (control group). The results for most of the determined enzymes revealed high activities as compared to the controls. The activity of acid phosphatase in male urine samples increased also, though not significantly. These elevated enzyme activities could be used to establish the diagnosis of schistosomiasis in patients whose urine contains no ova or when it is difficult to detect them. The results are discussed in the light of localization of each enzyme in the urinary tract as well as in other organs like the liver.

An immunocytochemical and morphometric study of the rat pancreatic islets.

The rat pancreas has frequently been used as an animal model to study changes in islet cells in pathological conditions, such as diabetes mellitus and islet cell tumours, but detailed quantitative data on the islets are not available. This study was therefore undertaken to investigate (1) the volume density of pancreatic islets, (2) islet diameter, islet volume and islet cell number and (3) islet cell pattern, i.e. the distribution, volume and number of each cell type per islet. The study also investigated the possibility of differences in various pancreatic regions derived from the dorsal primordium. The rat pancreas was divided into 4 regions: lower duodenal (derived from the ventral primordium) and upper duodenal, gastric and splenic regions (derived from the dorsal primordium). Sections were stained immunocytochemically with anti-insulin (B cells), antiglucagon (A cells), antisomatostatin (D cells) and antipancreatic polypeptide (PP cells) antibodies, and were used for morphometric analysis. A total of 1292 islets was examined, 328 from the lower duodenal, 245 from the upper duodenal, 314 from the gastric and 405 from the splenic regions. The mean volume density of the islets per pancreatic tissue was found to be 2.6 +/- 0.1%, 2.3 +/- 0.1%, 2.9 +/- 0.2% and 3.3 +/- 0.2%, in the lower duodenal, upper duodenal, gastric and splenic regions, respectively. The size-frequency distribution of the profile diameters of the islets showed an overall shift of all the size classes towards smaller sizes in the upper duodenal region, and towards larger sizes in the splenic region, as compared with the corresponding classes of the other regions.(ABSTRACT TRUNCATED AT 250 WORDS)

Marine algae that display anti-tumorigenic activity against Agrobacterium tumefaciens.

Thirty-five extracts representing different seasonal growths of 17 marine algal species collected from the Alexandria coast were tested for anti-tumorigenic activity against Agrobacterium tumefaciens galls on potato discs. Eleven extracts (nine species) displayed > 20% inhibition of tumor initiation, with three of these (Codium tomentosum, winter; Jania rubens, summer; Padina pavonia, winter) displaying relatively high activity. Bacterial viability tests showed that the inhibitory effects were directly due to anti-tumorigenesis rather than an indirect result of anti-bacterial activity.

Isolated pancreatic islets of the rat: an immunohistochemical and morphometric study.

Although there is a recent increase in the use of the isolated pancreatic islets of the rat in the transplantation and functional studies, there has been no detailed quantitative assessment on the size and cellular constituents of islets after the isolation procedure. The present work was undertaken to study the size classes of the isolated islets and the morphometry of their cellular populations. Islets of the rat pancreas were isolated by using the intraductal collagenase digestion technique, the most commonly used procedure for the isolation of pancreatic islets. Different endocrine cells of the isolated islets were stained by immunoperoxidase staining techniques. The distribution of the cellular constituents of the isolated islets was similar to that of the intact islets of the normal pancreas; A, D, and PP cells were peripherally arranged around the centrally located B cells. However, morphometric quantitative study showed that the percent volume and percent number of A, D, and PP cells of the isolated islets were lower than those of the corresponding intact ones. Further, the mean true diameter of the isolated islets was lower than that of the intact ones. These data indicate loss of islet cells during the process of isolation. Most of the lost cells were from the periphery of islets. This may provide an explanation for the incomplete metabolic control and recurrence of hyperglycemia encountered after isolated islet transplantation in the treatment of diabetes mellitus. It seems that further refinements of the isolation techniques are necessary to obtain islet tissue with total cellular integrity, before a complete success in transplantation could be achieved.

Ultrastructural observations on the hamuli of the monogenean gill parasite Cichlidogyrus halli typicus.

The ultrastructure of a monogenean hamulus is described for the first time. The ultrastructure of the dorsal and ventral hamuli of the monogenean gill parasite Cichlidogyrus halli typicus (Price & Kirk, 1967, 76: 137-144) Revue de Zoologie et de Botanique Africaines Paperna, 1979 (Annales du Musée Royal del'Afrique Centrale 226: 1-31) is similar but there are differences between different regions of a single hamulus. Each hamulus has three layers in the middle of the hook region, a thin outer layer, a middle layer forming the bulk of the hook and a central core made up of longitudinally oriented rods. The central core disappears in the distal region of the hook and the outer layer is absent in the proximal region of the hook. Both the shaft and the roots consist of two layers, the inner layer of the shaft is made up of longitudinally oriented parallel rods, and the corresponding layer of the roots being composed of long fibrils. The longitudinally orientated rods of the central core may provide inner strength to resist the physical strain on the hook and the outer, and possibly the middle layers, may provide chemical resistance to host's tissue secretion. The distal region of each root is covered with a fibrous hood, which appear to be a specialized region for binding muscle fibres to the sclerite.