A rare case and review of pulmonary ossification.

Pulmonary ossification (PO) is a rare metastatic disease characterized by the formation of diffuse heterotopic bone units in the lung parenchyma. Herein, we describe a 45-year-old Filipino male with dendriform pulmonary ossification in combination with gastroesophageal reflux disease and chronic dust exposure, a notably unique association. Radiographic imaging and pathology findings are examined with discussion of various pulmonary disease entities from current literature. Further recognition of PO will facilitate appropriate treatment and better outcomes for patients diagnosed with this enigmatic condition.

Light and transmission electron microscopy of the haptoral sclerites of the monogenean gill parasite Macrogyrodactylus clarii.

The present study represents the first detailed description of the haptoral sclerites of Macrogyrodactylus clarii Gussev 1961. Light microscopy reveals outgrowths of the hamuli roots; two lateral spine-like extensions of the hood-like accessory sclerites; a ridged, fan-shaped distal end of an accessory sclerite; and two thread-like accessory sclerites with biforked ends associated with the pointed hooked region of each hamulus. Transmission electron microscopy (TEM) reveals the presence of parallel tubules in the root of the hamuli and structural differences along the length of the hamulus including the root, shaft, and pointed hooked region. The root consists of two layers, the shaft four layers and pointed hooked region only one layer with dense outer serrations. Characteristic features of the hamulus root are the presence of longitudinally orientated parallel tubules in its central core and parallel electron-dense ridges in the outer layer of its middle region; features not observed in either the shaft or the pointed hooked region. Each hamulus blade of M. clarii is associated with haptoral gland cells producing electron-dense secretory bodies. The 16 marginal hooklets each consist of a blade (sickle) articulating with a handle at the guard region and a domus. TEM revealed structural differences between the handle, the blade at the articulation region, and the distal hooked region. The domus, a filamentous thread-like sclerite at the light microscope level, consists of two electron-dense, fibrous thickenings connected to each other by a cytoplasmic process. Each marginal hooklet is associated with a small cavity and a large reservoir of homogeneous particles and secretory bodies. The possible functions of these structures are discussed in relation to equivalent features in other monogeneans.

Ultrastructure of the anterior adhesive apparatus of the gill parasite Macrogyrodactylus clarii and skin parasite M. congolensis (Monogenea; Gyrodactylidae) from the catfish Clarias gariepinus.

Transmission electron microscopy (TEM) was used for the first time to study the anterior adhesive apparatus of the monogeneans Macrogyrodactylus clarii Gussev, 1961 and M. congolensis (Prudhoe, 1957) Yamaguti, 1963 inhabiting gills and skin respectively of the same catfish Clarias gariepinus. Despite the different microhabitats occupied by these parasites, the present study revealed that they have a similar anterior adhesive system. In both parasites, the anterior adhesive apparatus consists of three types of gland cells: G1 cells that produce rod-shaped bodies (S1), G2 cells manufacture irregularly shaped bodies (S2) and G3 cells form mucoid-like secretions (S3). In the cytoplasm of G1 cells, a single layer of microtubules encloses each developing rod-shaped body. A unique feature of S1 secretory bodies is that some fully developed S1 bodies are attached to each other, forming large condensed globules in the cytoplasm of G1 gland cells and terminal portion of the G1 ducts, but none were detected in the adhesive sacs outside the ducts. In the adhesive sacs, G1 ducts open with multiple apertures whereas each of the G2 and G3 ducts have a single opening. The adhesive sacs are lined with two types of tegument (st1 and st2). A third tegument type (st3) connects the st2 tegument with the general body tegument. Only st1 has microvilli. Each adhesive sac is provided with a spike-like sensillum and single uniciliated sense organ. The possible functions of microvilli in increasing the surface area and assistance in spreading and mixing of the adhesive secretion, and the role of sense organs associated with the adhesive sacs are discussed.

Scanning and transmission electron microscopy of the histopathological impact of Macrogyrodactylus clarii (Monogenea: Gyrodactylidae) on the gills of catfish, Clarias gariepinus.

Scanning and transmission electron microscopy (TEM) were used to study the histopathological effects of the monogenean Macrogyrodactylus clarii Gussev, 1961 on the gills of the catfish Clarias gariepinus (Burchell). Suction generated during attachment created 'footprints' on host surfaces in which the host tissues were elevated above the general gill surface. 'Footprints' were bordered by four clefts caused by the muscular flaps on the anterior, lateral and posterior margins of the haptor. The hamuli points penetrate the gill tissue but no evidence was found for the insertion of the marginal hooklets. At the site of attachment, host cells adjacent to the lateral flaps often appeared compressed and widely spaced with large intercellular spaces. Desquamation of these surface epithelia was also apparent and some of the widely spaced epithelial cells had pseudopodium-like processes. Cells within the upper surface epithelial layer of the host were vacuolated and necrotic. Ruptured blood capillaries (blood spaces) in the secondary gill lamellae contained atypical compressed erythrocytes, agranular and granular leucocytes and evidence of haemorrhaging. Cells with fibrotic cytoplasm, putative phagocytes and host mucous cells were evidence of a host response at the site of parasite attachment. The possible role of these cells is discussed in relation to host resistance against infection.

Regions of homozygosity identified by oligonucleotide SNP arrays: evaluating the incidence and clinical utility.

Copy neutral segments with allelic homozygosity, also known as regions of homozygosity (ROHs), are frequently identified in cases interrogated by oligonucleotide single-nucleotide polymorphism (oligo-SNP) microarrays. Presence of ROHs may be because of parental relatedness, chromosomal recombination or rearrangements and provides important clues regarding ancestral homozygosity, consanguinity or uniparental disomy. In this study of 14 574 consecutive cases, 832 (6%) were found to harbor one or more ROHs over 10 Mb, of which 651 cases (78%) had multiple ROHs, likely because of identity by descent (IBD), and 181 cases (22%) with ROHs involving a single chromosome. Parental relatedness was predicted to be first degree or closer in 5%, second in 9% and third in 19%. Of the 181 cases, 19 had ROHs for a whole chromosome revealing uniparental isodisomy (isoUPD). In all, 25 cases had significant ROHs involving a single chromosome; 5 cases were molecularly confirmed to have a mixed iso- and heteroUPD15 and 1 case each with segmental UPD9pat and segmental UPD22mat; 17 cases were suspected to have a mixed iso- and heteroUPD including 2 cases with small supernumerary marker and 2 cases with mosaic trisomy. For chromosome 15, 12 (92%) of 13 molecularly studied cases had either Prader-Willi or Angelman syndrome. Autosomal recessive disorders were confirmed in seven of nine cases from eight families because of the finding of suspected gene within a ROH. This study demonstrates that ROHs are much more frequent than previously recognized and often reflect parental relatedness, ascertain autosomal recessive diseases or unravel UPD in many cases.

On some ultrastructural features of the reproductive system of the monogenean parasite Macrogyrodactylus congolensis from Clarias gariepinus inhabiting the River Nile in Egypt.

Some organs of the reproductive system of the protogynous monogenean skin parasite Macrogyrodactylus congolensis (Prudhoe, 1957) Yamaguti, 1963 have been studied using transmission electron microscopy. The vesicula seminalis is enclosed by a prominent layer of circular muscle fibres and has inner syncytial protrusions. The penis bulb is a highly muscular organ with prominent radial and circular muscle fibres, a gutter-shaped large spine and 16 small spines. Two syncytial male accessory glands, and a single reservoir for male accessory secretion were identified. The secretory bodies in the male accessory glands and male accessory reservoir have a unique structure. A large oocyte is situated in a chamber, previously referred to as the "ootype" or "egg-cell-forming region" (ECFR), which also contains one or two small undifferentiated cells and vacuolated tissue. Mature spermatozoa were abundant in the receptaculum seminis and dispersed in the vacuolated tissue in the ECFR and appeared to be attached to the membrane of the large oocyte. Mature spermatozoa were also seen in the parenchymal tissue near the chamber containing embryos and even in the tissues of the embryo.

Ultrastructure of the digestive system and experimental study of feeding in the monogenean skin and fin parasite Macrogyrodactylus congolensis.

In the present study, transmission electron microscopy (TEM) has been used to study the ultrastructure of the digestive system, namely the pharynx, oesophageal glands and intestine, of the monogenean skin and fin parasite Macrogyrodactylus congolensis. The pharynx consists of an anterior highly muscular region and a posterior mainly glandular syncytial region. The anterior region is provided with six pharyngeal papillae, the centre of each of which is occupied by electron dense secretory bodies, identical with those in the posterior region of the pharynx. The intestine has an uninterrupted syncytial gastrodermis and the luminal surface is provided with many unbranched lamellae. The intestine of living specimens contains large and small granules which give it a reddish brown colour. Large particles, presumed to be lipid droplets, and small granules, presumed to be melanin granules, were found in the gastrodermis and in the intestinal lumen. Parasites were induced to feed and then preserved for TEM at the following intervals: just after feeding, 30 min after feeding, 1 h 30 min after feeding and 2 h after feeding. The specimens were then processed for TEM and sections cut through the intestine of each specimen were examined with the transmission electron microscope. Three types of vacuoles (V1, V2, V3) were detected in the gastrodermis. Vacuoles V1 have thick walls and are likely to be endocytotic, enclosing luminal contents at the surface of the gastrodermis. V2 vacuoles may be lysosomes that fuse with V1 vacuoles. V3 vacuoles may serve to dispose of residual digestive material into the lumen.

Genotype-phenotype analysis of recombinant chromosome 4 syndrome: an array-CGH study and literature review.

BACKGROUND: Recombinant chromosome 4, a rare constitutional rearrangement arising from pericentric inversion, comprises a duplicated segment of 4p13~p15→4pter and a deleted segment of 4q35→4qter. To date, 10 cases of recombinant chromosome 4 have been reported. RESULT: We describe the second case in which array-CGH was used to characterize recombinant chromosome 4 syndrome. The patient was a one-year old boy with consistent clinical features. Conventional cytogenetics and FISH documented a recombinant chromosome 4, derived from a paternal pericentric inversion, leading to partial trisomy 4p and partial monosomy of 4q. Array-CGH, performed to further characterize the rearranged chromosome 4 and delineate the breakpoints, documented a small (4.36 Mb) 4q35.1 terminal deletion and a large (23.81 Mb) 4p15.1 terminal duplication. Genotype-phenotype analysis of 10 previously reported cases and the present case indicated relatively consistent clinical features and breakpoints. This consistency was more evident in our case and another characterized by array-CGH, where both showed the common breakpoints of p15.1 and q35.1. A genotype-phenotype correlation study between rec(4), dup(4p), and del(4q) syndromes revealed that urogenital and cardiac defects are probably due to the deletion of 4q whereas the other clinical features are likely due to 4p duplication. CONCLUSION: Our findings support that the clinical features of patients with rec(4) are relatively consistent and specific to the regions of duplication or deletion. Recombinant chromosome 4 syndrome thus appears to be a discrete entity that can be suspected on the basis of clinical features or specific deleted and duplicated chromosomal regions.

Neocentric X-chromosome in a girl with Turner-like syndrome.

BACKGROUND: Neocentromeres are rare human chromosomal aberrations in which a new centromere has formed in a previously non-centromeric location. We report the finding of a structurally abnormal X chromosome with a neocentromere in a 15-year-old girl with clinical features suggestive of Turner syndrome, including short stature and primary amenorrhea. RESULT: G-banded chromosome analysis revealed a mosaic female karyotype involving two abnormal cell lines. One cell line (84% of analyzed metaphases) had a structurally abnormal X chromosome (duplication of the long arm and deletion of the short arm) and a normal X chromosome. The other cell line (16% of cells) exhibited monosomy X. C-banding studies were negative for the abnormal X chromosome. FISH analysis revealed lack of hybridization of the abnormal X chromosome with both the X centromere-specific probe and the "all human centromeres" probe, a pattern consistent with lack of the X chromosome endogenous centromere. A FISH study using an XIST gene probe revealed the presence of two XIST genes, one on each long arm of the iso(Xq), required for inactivation of the abnormal X chromosome. R-banding also demonstrated inactivation of the abnormal X chromosome. An assay for centromeric protein C (CENP-C) was positive on both the normal and the abnormal X chromosomes. The position of CENP-C in the abnormal X chromosome defined a neocentromere, which explains its mitotic stability. The karyotype is thus designated as 46,X,neo(X)(qter- > q12::q12- > q21.2- > neo- > q21.2- > qter)[42]/45,X[8], which is consistent with stigmata of Turner syndrome. The mother of this patient has a normal karyotype; however, the father was not available for study. CONCLUSION: To our knowledge, this is the first case of mosaic Turner syndrome involving an analphoid iso(Xq) chromosome with a proven neocentromere among 90 previously described cases with a proven neocentromere.

Association of the CagA gene positive Helicobacter pylori and tissue levels of interleukin-17 and interleukin-8 in gastric ulcer patients.

It has been reported that CagA gene positive Helicobacter pylori (CagA+ H. pylon) induces severe gastric mucosal inflammation. On the other hand, Interleukin (IL)-17 is known to stimulate IL-8 release by the gastric epithelial cells which facilitates chemotaxis of neutrophils through an IL-8-dependent mechanism. The aim of the study is to determine the role of IL-17 and IL-8 in the development of gastritis and gastric ulcer in H. pylori infected patients. Mucosal biopsy samples were obtained from the ulcer site of gastric mucosa of 28 patients with gastric ulcer (GU), 27 with gastritis and 8 controls subjects without gastritis or ulcers. Infection with H. pylori of patients and controls was assessed by a rapid urease test, histological examination and culture. Measurement of the tissue levels of IL-17 and IL-8 were assayed by ELISA. H. pylori cagA gene was assessed by polymerase chain reaction (PCR). Out of the 28 patients with GU, 18 (64.2%) patients were positive for H. pylori infection, while 13 (48.1%) patients with gastritis and none of the controls were positive for H. pylori infection The CagA gene was detected in 12 (66.6%) in H. pylori GU patients, and 7 (53.8%) H. pylori positive gastritis. IL-17 was significantly higher in GU-CagA+ve H. pylori compared to GU-CagA- H. pylori (P <0.05), while IL-8 showed no significant difference between groups. The mean levels of IL-8 in gastritis-CagA+ H. pylori) was significantly higher compared to gastritis--CagA- H. pylori- (P <0.05). IL-17 showed significant association with the number of neutrophils in both GU and gastritis (r = 0.689, P < 0.05 & r = 0.618, P < 0.05). Also, IL-8 showed significant association with the number of neutrophils in both GU and gastritis n (r = 0.468, P < 0.05 & r = 0.727, P < 0.05). It is concluded that the Cag+ve H. pylori is associated with induction of mucosal injury. Also, IL-8 and IL-17 plays a role in the development of GU and gastritis especially in CagA+ H. pylori.

Sinus aspirates in chronic rhinosinusitis: fungal colonization of paranasal sinuses, evaluation of ICAM-1 and IL-8 and studying of immunological effect of long-term macrolide therapy.

PURPOSE: In patients with chronic fungal sinusitis, concentrations of interleukin-8 (IL-8), immunoglobulin E (IgE), and soluble intercellular adhesion molecule-1 (sICAM-1) were compared in paranasal sinus aspirates and serum. Furthermore, immunological effects of macrolide treatment of our patients with chronic fungal rhinosinusitis were also studied. MATERIAL AND METHODS: In our cohort study, 108 patients with chronic rhinosinusitis undergoing sinus surgery were selected. Sinus aspirates were collected, and used for immunological assasy and cultured for fungal study. All patients were examined for the presence of characteristic allergic mucin of chronic allergic fungal rhinosinusitis and this was confirmed later by measurement of total serum IgE. RESULTS: Our cases were classified into 3 groups: chronic rhinosinusitis with positive fungal culture and negative allergic mucin, chronic rhinosinusitis with positive fungal culture and positive allergic mucin and chronic rhinosinusitis without fungal growth. A control group was included. We found 57.4% of the patient cultures positive for fungus and 36.4% of the control subjects. Aspergillus ssp. were the most prevalent followed by Bipolaris ssp., and Curvularia. IgE levels were increased in group II compared to group I, III and IV. ICAM-1 and IL-8 levels were increased in groups I, II and III compared to the control group. Erythromycin given in group II decreased the levels of IL-8 and ICAM-1. CONCLUSION: Aspergillus species were the most common. These results confirm the role of ICAM-1 and IL-8 in all types of rhinosinusitis. Erythromycin modulated the immune status of the patients.

Bacterial DNA and its consequences in patients with cirrhosis and culture-negative, non-neutrocytic ascites.

The detection of bacterial DNA in serum and ascitic fluid (AF) from patients with liver cirrhosis and ascites is interpreted as molecular evidence of intestinal bacterial translocation (BT) and considered sufficient to activate the cellular immune response leading to greater cytokine synthesis. We studied 34 patients with liver cirrhosis and culture-negative, non-neutrocytic ascites [22 patients without bacterial DNA (group I) and 12 patients with bacterial DNA (group II)]. History and clinical examination were done with the following investigations at first admission and followed up for 24 weeks: serum and AF tumour necrosis factor-alpha (TNF-alpha), AF polymorphonuclear leukocytes, AF cultivation and detection of blood and AF bacterial DNA. Serum and AF TNF-alpha were significantly higher in patients with bacterial DNA compared to those without bacterial DNA at first admission [54.5+/-22.56 vs 35.2+/-17.97 pg ml(-1) (P=0.02) and 123.2+/-49.32 vs 82.6+/-29.58 pg ml(-1) (P <0.005), respectively]. These changes became highly significant at the end of follow-up of both groups [119.3+/-27.19 vs 40.2+/-16.08 pg ml(-1) (P <0.001) and 518.8+/-91.11 vs 97.6+/-17.81 pg ml(-1) (P <0.001), respectively]. In group II, there was a significant increase in serum and AF TNF-alpha at the end of follow-up compared to at first admission (P <0.001). The relative risk of death, hepatorenal syndrome (HRS) and spontaneous bacterial peritonitis (SBP) was higher in patients with bacterial DNA compared to those without bacterial DNA. We conclude that cirrhotic patients with culture-negative, non-neutrocytic ascites and bacterial DNA have a significantly higher level of serum and AF TNF-alpha and higher risk of HRS, SBP and mortality compared to those without bacterial DNA, suggesting that bacterial DNA and TNF-alpha are implicated in these complications of liver cirrhosis.

Neuromusculature of Gyrodactylus rysavyi, a monogenean gill and skin parasite of the catfish Clarias gariepinus.

Phalloidin fluorescence technique, enzyme cytochemistry and immunocytochemistry in conjunction with confocal scanning laser microscopy were used for the first time to describe the nervous and muscle systems of the viviparous monogenean parasite, Gyrodactylus rysavyi inhabiting the gills and skin of the Nile catfish Clarias gariepinus. The body wall muscles are composed of an outer layer of 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 musculature of the pharynx, intestine, reproductive tract and the most prominent muscles of the haptor were also described. Two characteristic muscular pads were found lying in the anterior region of the haptor in close contact with the hamuli. To each one of these pads, a group of ventral extrinsic muscles was connected. The role of this ventral extrinsic muscle in the body movement was discussed. The mechanism operating the marginal hooklets was also discussed. The central nervous system (CNS) 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. The CNS is better developed ventrally than dorsally or laterally and it has the highest reactivity for all neuroactive substances examined. Both the central and the peripheral nervous system (PNS) are bilaterally symmetrical. Structural and functional correlates of the neuromusculature of the pharynx, haptor and reproductive tracts were explained. The results implicated acetylcholine, FMRFamide-related peptides (FaRPs) and serotonin in sensory and motor function. The results were compared with those of the monogeneans Macrogyrodactylus clarii and M. congolensis inhabiting the gills and skin respectively of the same host fish C. gariepinus.