Naked DNA immunization with full-length attachment gene of human respiratory syncytial virus induces safe and protective immune response.

Development of potent vaccine for human respiratory syncytial virus (HRSV) that confers better protection than natural infection remains a global challenge. Vaccination with naked DNA is considered successful approach for the control of many viral diseases. In this study, the potential of DNA vaccination using full-length attachment gene of HRSV type A Saudi strain cloned in pcDNA3.1+ vector (pcDNA/GA) was evaluated in BALB/c mice. The expression efficiency of pcDNA/GA was first confirmed in HEp-2 cells on RNA and protein levels. Mice immunization with either pcDNA/GA or the positive control formalin-inactivated vaccine (FI-RSV) has generated significant serum antibody concentration in ELISA (7.31±0.418 and 9.76±0.006 µg/ml, respectively) with superior neutralizing activity. Similarly, both immunogens evoked robust HRSV-specific CD8+ T-cell response in ELISPOT assay compared to mice immunized with pcDNA3.1+ vector or saline (negative controls). Challenge of the immunized mice with the wild-type HRSV did not provoke clinical symptoms or mortality in any mice group. On the 7th day post-challenge, mice were euthanized and lungs were extirpated for evaluation of viral load, histopathological changes and cytokine profile. A significant diminish in the viral load and histology score were concluded in lungs of pcDNA/GA immunized mice compared to those immunized with FI-RSV and negative controls. The pulmonary cytokine profile of pcDNA/GA immunized mice displayed notable upregulation of Th1-associated cytokines while that of FI-RSV immunized mice exhibited high levels of Th2-associated cytokines. In conclusion, the DNA vaccine candidate pcDNA/GA has proven prominent efficacy and safety in mouse model, which encourages further evaluation in clinical trials.

[Psychiatric care of refugees in Africa and the Middle East : Challenges and solutions]. / Psychiatrische Versorgung von Flüchtlingen in Afrika und dem Nahen Osten : Herausforderungen und Lösungsansätze.

Violence, flight, famine, and natural disasters as well as the absence of a psychosocial healthcare system are major psychological burdens for refugees. The level of provision of mental healthcare is particularly low in developing countries. Internally displaced people and refugees place high demands on the healthcare system because they often suffer from psychiatric disorders, such as depression, posttraumatic stress disorder, and substance use disorders. We present first initiatives to improve psychiatric care in refugee camps in Ethiopia, Kenya, and Sudan. Moreover, we provide first insights into a project based in Northern Iraq and Germany aimed at the treatment of people who were severely traumatized by the terror regime of the so-called Islamic State (IS).

Prenatal metanephrogenesis of the camel: morphological evidence of epithelial-mesenchymal interaction.

The present investigation examined histogenesis of epithelial, stromal and angiogenic elements of the prenatal camel permanent or metanephric kidney. The primitive metanephros was first observed at the 13-mm crown vertebral rump length (CVRL) stage as an ovoid structure composed of a centrally located epithelial ureteric bud and peripheral circumscribed masses of undifferentiated mesenchymal cells. The first morphological evidence of glomerulogenesis was observed at the 28-mm CVRL stage. Developing renal corpuscles became obvious at the 35-mm CVRL stage. At the 60-mm CVRL stage, the epithelial renal pelvis gave rise to tubular branches that extended towards the cortical zone. These branches represented the presumptive collecting ducts. Differentiation of renal tubules into the proximal and distal convoluted tubules was observed at the 95-mm CVRL stage. At the 130-mm CVRL stage, the renal medulla was clearly delineated into medullary pyramids, which in association with the corresponding cortical caps formed the morphological basis of the renal lobar formation. A gradual nephrogenic decline was noticed from the 940-mm CVRL on; however, the process of nephrogenesis persisted throughout all the studied foetal stages.

Apoptosis in the myocardium of the adult dromedary camel: ultrastructural characterization.

Apoptosis is a highly regulated mode of cell death that occurs in the absence of inflammation. Light microscopic (LM) examination of the myocardium of apparently healthy camel did not reveal evidence of apoptosis in any samples; however, evidence of apoptosis was apparent by transmission electron microscopy (TEM). The most common apoptotic features observed by TEM included (1) an intact sarcolemma with some bleb formation; (2) nuclear chromatin condensation and margination with nucleolar disruption; (3) mitochondrial swelling and disorganization, accompanied by degeneration or hypercondensation of cristae; and (4) an intercalated disc region with a higher-than-normal mitochondrion/myofibril ratio, or surrounded from both sides by asymmetrically contracted sarcomeres. Apoptotic alterations were also noted among the endothelial cells lining the microvasculature of the myocardium. These alterations included (1) marked nuclear chromatin condensation and margination; (2) villous blebs on the adluminal plasmalemma, which projected into the lumen; (3) cytoplasmic vacuolation; (4) presence of intraluminal membrane-bounded vesicles; and (5) occasional pericapillary edema and accumulations of cellular debris. The results of this study indicate that myocardial apoptosis can occur in apparently healthy camels, in the absence of a clear-cut etiology.

Immunohistochemical mapping of natriuretic peptide receptor-A in the brainstem of Macaca fascicularis.

Natriuretic peptide receptor-A (NPR-A) mediates the biological effects of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), and is involved in maintaining cardiovascular homeostasis. In this immunohistochemical study we examined the distribution of NPR-A in the brainstem of the cynomolgus monkey. NPR-A immunoreactivity was localized to neurons in specific brainstem regions. NPR-A-immunoreactive perikarya were found in the red nucleus and the oculomotor nucleus in the midbrain, the parabrachial nucleus and the locus coeruleus in the pons, and the dorsal motor nucleus of the vagus, the hypoglossal nucleus, the cuneate nucleus, the gracile nucleus, the nucleus ambiguus, the lateral reticular nucleus, the reticular formation, and the inferior olivary nucleus in the medulla oblongata. Extensive networks of immunoreactive fibers were apparent in the red nucleus, the oculomotor nucleus, the principal sensory trigeminal nucleus, and the parabrachial nucleus. Double immunostaining revealed NPR-A immunoreactivity in cholinergic neurons of the parabrachial nucleus, the dorsal motor nucleus of vagus, the hypoglossal nucleus, and the nucleus ambiguus. However, there was no colocalization of NPR-A and tyrosine hydroxylase in the locus coeruleus. The wide anatomical distribution of NPR-A-immunoreactive structures suggests that natriuretic peptides, besides having a role in the central regulation of endocrine and cardiovascular homeostasis, may also mediate diverse physiological functions.

Ultrastructural analysis of the integument of a desert-adapted mammal, the one-humped camel (Camelus dromedarius).

In this study, we conducted a light microscopic and ultrastructural analysis of the integument of the one-humped camel (Camelus dromedarius). In general, the epidermal strata of the camel integument appeared typical of those found in non-desert mammals. Two cell populations were noted in the stratum basale: one with a flat, non-serrated base and the other with a highly serrated base. Typical fine structure was observed in keratinocytes of the stratum spinosum and stratum granulosum. The stratum corneum was six to 10 cells thick. Within the different strata, overall cell morphologies and the general distribution and relative abundance of cellular organelles appeared typical. Dermal features included the presence of myoepithelial cells surrounding apocrine tubular glands. Inter- or intracellular canaliculi within the secretory cells of the apocrine glands, reported to be present in certain other non-desert mammals, were not evident in the camel. Together, these data indicate that while the camel is clearly adapted for a desert lifestyle, these adaptations do not include significant specializations at the cellular or subcellular level in the integument.

Voltammetric analysis of Cu (II), Cd (II) and Zn (II) complexes and their cyclic voltammetry with several cephalosporin antibiotics.

Both osteryoung square wave voltammetry and cyclic voltammetry have been utilized to elucidate and confirm the possible complexation reaction that occur between the various cephalosporin antibiotics and either the toxic, non-essential metal ion, viz. Cd (II), or the essential but toxic (when their concentration exceeds certain level in serum) metal ions, viz. Cu (II) and Zn (II). Voltammetric measurements indicated the existence of 1:1 metal-to-ligand ratio (as in cephalexin and cephapirin complexes), 1:2 ratio (such as in cefamandole, cefuroxime and cefotaxime complexes) and 2:1 ratio in case of ceftazidime complexes. Adsorption behavior was evidenced for Cu (II)-cefuroxime or ceftazidime complexes as well as for those for Zn (II)-cephalexin or cephapirin. This phenomenon could be used for the determination of either the antibiotic or the metal ion using adsorptive stripping voltammetry. Detection limits down to 7x10(-10) M have been easily achieved.

Molecular identification and immunohistochemical localization of atrial natriuretic peptide in the heart of the dromedary camel (Camelus dromedarius).

Atrial and B-type natriuretic peptide (ANP and BNP) are cardiac hormones synthesized and secreted by the myoendocrine cells of the heart. They exert potent actions on body fluid balance. Since various body organs including the heart are under high physiological stress during water and food deprivation in the desert nomads, we intended to perform molecular biological and histological studies of ANP in the heart of the dromedary camel Camelus dromedarius. Initially, we isolated cDNAs encoding ANP from the atrium and BNP from the atrium and ventricle of the dromedary camel. Putative mature ANP, deduced from the cDNA sequence, was identical to that of human and pig ANP, but the putative mature BNP was more diverse and was most similar to pig BNP (94% identity). Thus, we used antisera raised against human ANP that did not cross-react with pig BNP in the subsequent immunohistochemical studies. The ANP-expressing myoendocrine cells are most concentrated in the right atrium, to a lesser extent in the left atrium, and almost absent in the left ventricle. The immuno-positive cells are scattered uniformly in each region and are characterized by the presence of immunoreactive granular deposits around the nucleus. The left atrium comprises some ramifications of conductive cells (Purkinje fibers), some of which also contained ANP-immunoreactive granules. At the electron microscopic level, myoendocrine cells possessed secretory granules primarily in the perinuclear zone and a well-developed Golgi apparatus. The present study is the first comprehensive report dealing with the molecular cloning and immunohistochemical localization of ANP in the heart of a desert dwelling mammal.

Immunolocalization of relaxin in the corpus luteum of the dromedary camel. (Camelus dromedarius).

The objective of this study was to determine the distribution of the peptide hormone relaxin in ovarian tissues of the pregnant and non-pregnant dromedary (Camelus dromedarius). Using light and electron microscopy we here describe the presence of relaxin in cells of the corpus luteum at various stages of the reproductive cycle of this important species.

Liver ultrastructure and a new cell type in the Japanese newt, Cynops pyrrhogaster.

The liver of the Japanese newt, Cynops pyrrhogaster, has been investigated using light, scanning, and transmission electron microscopy. Hepatic parenchyma was composed of clusters and cords or tubules of polyhedral cells separated by a sinusoidal net. Hepatocytes had spherical, euchromatic nuclei with one or more nucleoli and stacked mitochondria with sparse cristae and dense bodies. Rough endoplasmic reticula formed peribiliary stacks and diffusely scattered vesicles and tubules. Smooth endoplasmic reticula were more pronounced in glycogen-rich hepatocytes. Most hepatocytes contained peroxisomes, Golgi complexes and large numbers of fat droplets within the cytoplasm along with glycogen. Some cells were mainly glycogen-storing and contained few or no fat droplets. A special feature of the newt liver was biliary atresia. Bile canaliculi had short, stout microvilli which were entirely atretic in some canaliculi. Canaliculi were sealed off by junctional complexes including zonulae occludentes and maculae adherentes. The latter showed extraordinary wider desmosomal gaps in the vicinity of the atretic bile canaliculi. The sinusoid wall was non-distinctive and contained fenestrated endothelial cells connected to Kupffer cells by zonulae occludentes. A distinctive new cell type (OG cell) was observed in the newt liver. These cells were found individually or in small clusters in proximity with the sinusoidal surfaces. They had small nuclei, a paucity of cytoplasmic organelles, but numerous, unique, osmiophilic granules of two distinct types. Less numerous Type I granules contained homogeneous electron-dense material, and a predominant Type II granule contained circumferentially arranged subparticulation. Granules of both types were detected within the cytoplasm of endothelial cells and within sinusoids together with blood elements. The function of this secretory type cell remains obscure, though it may represent a stage of melanophore.

Studies on the histogenesis of the tunica mucosa of the stomach of the Egyptian water buffalo (Bos bubalus L.). V. Histogenesis of the abomasal mucosa.

The primordia of the mucosal longitudinal folds are already prominent in the fetus with 9 cm CRL. In the fetus with 9 cm CRL the epithelium is stratified and becomes mixtured of simple and pseudostratified columnar in the fetus with 16 cm CRL in which the primordia of the abomasal glands are firstly seen but finally in the fetus with 17 cm CRL the whole epithelium becomes simple columnar. Although some few parietal cells are evident in the fetus with 73 cm among some fundic glands, but the complete-establishment of their cellular elements is completed postnatally. The Lamina muscularis mucosae is firstly seen in the fetus with 16 cm in the terminal portion of the abomasum.

Studies on the histogenesis of the tunica mucosa of the small and large intestines of the camel (Camelus dromedarius). II. Histogenesis of the colonic mucosa.

The colon is at first a simple tube, and its mucosa has a smooth luminal surface, surrounding a previous lumen. At the 150 mm stage the mucosa is thrown into a number of stout projections, each has a vascularized mesenchymal core and an epithelial cloth. These projections are in turn split into finger-like villi by transverse furrows. The villi reach their maximal length at the 260 mm stage and then gradually degenerate and slough into the lumen. The villi were completely disappeared at the 730 mm stage. The colonic glands were firstly observed at the 200 mm stage as downgrowths of the intervillous epithelium and they increase in number by dichotomous division of their bases. The Lamina muscularis mucosae is completely constructed at the 730 mm stage.

Studies on the histogenesis of the tunica mucosa of the small and large intestines of the camel (Camelus dromedarius). I. Histogenesis of the small intestinal mucosa.

The histogenesis of the small intestinal mucosa passes through a previllous stage, comprising development until the formation of villi (at a C RL of about 15 cm), and a villous stage, comprising the further development. In the previllous stage, the mucosa consists of a Lamina epithelialis made up of undifferentiated cells with stratified nuclei and a Lamina propria/submucosa made up of homogeneous mesenchyme. The villi appear as luminal epithelial outgrowths associated with vascularized mesenchymal stalks and their growth advances in a cranio-caudal direction. The intestinal glands were firstly observed at the 26 cm-stage as downgrowths of the intervillous epithelium into the underlying mesenchyme.

Studies on the histogenesis of the tunica mucosa of the stomach of the Egyptian water buffalo (Bos bubalus L.) III. Histogenesis of the mucosa of the ventricular sulcus (esophageal groove).

In the fetus with 9 cm CRL the ventricular groove was made up of two lips and a floor. The mucosa consisted of a Lamina epithelialis and a Lamina propria mucosae. The basal zone of the Lamina epithelialis was uneven in the floor of the groove and showed wedge-shaped evaginations into the superficial one. The luminal surface of the Lamina epithelialis of the floor became undulant in the fetus with 27 cm CRL. The Lamina muscularis mucosae became evident in the fetus with 73 cm CRL. In this fetal stage the Lamina epithelialis of the floor became differentiated into the Strata basale, spinosum, granulosum and corneum.

Studies on the histogenesis of the Tunica mucosa of the stomach of the Egyptian water buffalo (Bos bubalus L.). IV. Histogenesis of the omasal mucosa.

In the fetal stage of 9 cm CRL 4 order omasal laminae are present. With increasing age they increase greatly in size and conical papillae are seen projecting from their sides into the lumen. At 27 cm CRL stage sporadic smooth muscle cells in the interlaminar spaces, representing the first appearance of the Lamina muscularis mucosae, become visible in the cores of the laminae in the fetus with 32 cm CRL. A fifth order lamina occurs in the fetus with 32 cm CRL as evaginations of epithelium and the underlying Lamina propria mucosae in the interlaminar spaces. The epithelium becomes stratified squamous keratinized assuming the features of the ruminal epithelium after birth.

Studies on the histogenesis of the tunica mucosa of the stomach of the Egyptian water buffalo (Bos bubalus L.). II. Histogenesis of the reticular mucosa.

In the fetus with 9 cm CRL the reticular mucosa was made up of a Lamina epithelialis and a Lamina Propria/submucosa. The primordia of the reticular crests firstly appeared in the fetus with 16 cm CRL as evaginations of the basal zone of the Lamina epithelialis into the superficial one. Invaginations from the luminal surface of the Lamina epithelialis above the tips of the long reticular crests were formed in the fetus with 27 cm CRL. Aggregations of smooth muscle cells appeared in the cores of the long reticular crests in the fetus with 50 cm CRL. In the fetus with 73 cm CRL invaginations of various depths were formed to separate the reticular crests from the mass of epithelium. The epithelium in the different fetal stages were missing a Stratum corneum.

Studies on the histogenesis of the tunica mucosa of the stomach of the Egyptian water buffalo (Bos bubalus L.).

The histogenesis of the ruminal mucosa was studied in 9 buffalo fetuses (CRL 90--730 mm) and one suckling female buffalo calf. The Lamina epithelialis was found to consist of a basal and superficial layer. The former consists firstly of 2--4 cell layers and becomes later reduced to only one, made up to columnar cells; the latter one consists firstly of about 9 cell layers and increases then to 28--35, depending on the ruminal compartment. The epithelium shows its embryological feature, i.e. all nuclei are directed to the luminal cell pole. The first incidence of the ruminal papillary formation in the Egyptian water buffalo is observed in fetuses of 170 mm CRL. The histogenetic steps of the formation of the ruminal papillae are the aggregation of the cells of the basal layer and of the Lamina propria; the undulations with involvement of the basal layer of the Lamina epithelialis, basement membrane and Lamina propria; formation of humps from the undulations; formation of papillae. The papillary primordia are seen first in the Atrium ruminis and in the caudoventral blind sac simultaneously. The suckling calf has still no definite ruminal papillae. Only their tips are projecting to a different extent into the lumen.