Phe354Leu polymorphism of the liver kinase B1 gene as a prognostic factor in adult egyptian patients with acute myeloid leukemia.

Background: The human liver kinase B1 (LKB1) gene is a significant tumor suppressor widely expressed in all fetal and adult tissues. Despite its established role in solid tumors, the biological and clinical implications of LKB1 gene alterations in hematological malignancies have not been sufficiently recognized. Aim: This study aimed to determine the frequency of the LKB1 Phe354Leu polymorphism in adult Egyptian patients with cytogenetically normal AML (CN-AML), evaluate its clinical prognostic significance, and investigate its effect on the therapeutic outcome and patient survival. Methods: Direct sequencing of amplified exon eight of the LKB1 gene was performed to detect the Phe354Leu polymorphism in 72 adult de novo CN-AML patients. Results: The LKB1 Phe354Leu polymorphism was detected in 16.7% of patients and associated with younger age and lower hemoglobin levels (p < 0.001). Patients in the mutated group had significantly higher total leukocytic count and bone marrow blasts (p = 0.001 and p < 0.001, respectively). The most common FAB subtypes in mutated patients were M4 and M2. The relapse rate was significantly higher in the mutated group (p = 0.004). There was a significant association between the FLT3-ITD polymorphism and LKB1 F354L (p < 0.001). The mutated group had shorter overall survival (p = 0.003). In multivariate analysis, the Phe354Leu polymorphism was a significant independent prognostic variable for the overall and disease-free survival of the studied patients (p = 0.049). Conclusion: The LKB1 Phe354Leu polymorphism was diagnosed at younger ages in Egyptian CN-AML patients and represented a poor independent prognostic factor in CN-AML. Patients who carried this polymorphism had shorter overall survival and more frequent relapses. Our findings may provide insight into the design of therapeutic targets, and molecular testing of the LKB1 gene is recommended for proper risk stratification of CN-AML patients.

Evaluation of hazards of electronic -cigarette's liquid refill on testes of mice, complemented by histopathological and chromatographic analysis.

Electronic-cigarettes (e-cigarettes) are devices designed to become an alternative to classic cigarettes. Vaping of e-cigarettes and their recharge liquid have become extremely popular among the adolescents; however, its safety is not well established. Evaluation of the components of e-cigarette liquid and their potential effects on testis of adult male mice. This aim will be fulfilled by histological, ultrastructural, and immunohistochemical analysis of mice testis biopsies. Twenty mice were allocated into two groups of equal size. The control group was given regular saline, whereas the treated group was given e-liquid (contains 3 mg of nicotine/kg of body weight) both groups daily intraperitoneally injected for 3 weeks. Analysis of e-liquid by Gas Chromatography-Mass Spectrometric GC/MS demonstrated nicotine, phenol, vanillin, aldehydes, and pyrethroid insecticide. Evaluation of oxidative stress parameters revealed significant reduction of SOD and GPx. Histological results revealed a significant reduction in the height of seminiferous tubules, sloughing of spermatogenic cells, most cells being dark and pyknotic, and thickening of the interstitium with accumulation of PAS positive exudate. Most spermatogenic cells showed degenerative changes as rarefied cytoplasm, ill-defined electron-dense nuclei, and elongated spermatid showed deformity of ectoplasmic specialization. Immunohistochemical studies revealed a significant increase in caspase-3 positive cells and a significant reduction of area % of E-cadherin. The analysis of an available E-liquid demonstrated potentially harmful chemicals that are not shown in the labeling of the product. E-liquid appears to impair anti-oxidant defense and cause degenerative changes in the body and disruption of blood testes barrier BTB. So, e-cigarettes cannot be regarded as a non-harmful smoking replacement.

Testing efficacy of the nicotine protection of the substantia nigra pars compacta in a rat Parkinson disease model. Ultrastructure study.

Parkinson's disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) with subsequent motor manifestations. This study aimed to assess the ameliorative effects of nicotine, in rotenone-induced PD rat model. Thirty adult male Albino Wistar rats were divided into three equal groups. Group I received an injection of normal saline. Group II received subcutaneous injection of rotenone at a dose of 1.5 mg/kg every other day. Group III received rotenone in the same previous dose and nicotine at a dose of 1.5 mg/kg daily. After 11 days of treatment, body weight (BW) and rat motor behavior were estimated. Specimens from the midbrain were processed for light and electron microscopy. The expression of tyrosine hydroxylase (TH), α-synuclein, and GFAP was examined. Serum levels of total antioxidant capacity (TAC) and malondialdehyde (MDA), and striatal levels of dopamine (DA) were analyzed. Group III revealed a significant improvement in BW and motor activity. Nicotine upregulated the expression of TH, downregulated the expression of α-synuclein and GFAP. The levels of MDA and TAC were improved but were still far from those of the control. Striatal DA levels increased. Nicotine activated the neurons and glial cells. The vascular endothelium, however, did not elicit improvement. Although nicotine ameliorated the loss of the dopaminergic neurons and motor deficit, it did not show improvement of vascular endothelium. It is still necessary to examine nicotin's ability to maintain the dopaminergic neurons in a good functioning state.

Naproxen as a potential candidate for promoting rivastigmine anti-Alzheimer activity against aluminum chloride-prompted Alzheimer's-like disease in rats; neurogenesis and apoptosis modulation as a possible underlying mechanism.

BACKGROUND AND AIM: Alzheimer's disease (AD) is one of the leading causes of dependence and disability among the elderly worldwide. The traditional anti-Alzheimer medication, rivastigmine, one of the cholinesterase inhibitors (ChEIs), fails to achieve a definitive cure. We tested the hypothesis that naproxen administration to the rivastigmine-treated aluminum chloride (AlCl3) Alzheimer's rat model could provide an additive neuroprotective effect compared to rivastigmine alone. MATERIALS AND METHODS: The studied groups were control (Cont), AlCl3 treated (Al), rivastigmine treated (RIVA), naproxen treated (Napro), and combined rivastigmine and naproxen treated (RIVA + Napro). Rats' memory, spatial learning, and cognitive behavior were assessed followed by evaluation of hippocampal acetylcholinesterase (AChE) activity. Hippocampal and cerebellar histopathology were thoroughly examined. Activated caspase-3 and the neuroepithelial stem cells marker; nestin expressions were immunohistochemically assayed. RESULTS: AD rats displayed significantly impaired memory and cognitive function, augmented hippocampal AChE activity; massive neurodegeneration associated with enhanced astrogliosis, apoptosis, and impaired neurogenesis. Except for the enhancement of neurogenesis and suppression of apoptosis, the combination therapy had no additional neuroprotective benefit over rivastigmine-only therapy. CONCLUSION: Naproxen's efficacy was established by its ability to function at the cellular level, improved neurogenesis, and decreased, apoptosis without having an additional mitigating impact on cognitive impairment in rivastigmine-treated AD rats.

Nicotine impact on rat substantia nigra compacta.

Nicotine neuronal interactions exert an adverse potential in some brain regions and a significant link has been established between tobacco smoke/nicotine and vascular impairment. This work addresses nicotine impact on various components of the substantia nigra compacta (SNc) in rat. Twenty adult male Albino rats were divided equally into two groups: Group I, vehicle-control group (received saline [1 ml/kg body weight intra peritoneally] for 11 days). Group II; nicotine group (received 1.5 mg/kg body weight/day Sc) for 11 days. Nicotine levels were detected in the serum. Specimens were taken from the mid brain, processed and examined using biochemical, immunohistochemical, ultrastructural and morphometric techniques. In nicotine group, biochemical analysis revealed reduction in total antioxidant capacity (TAC), decrease in dopamine and malondialdehyde (MDA) levels. The mean number of light cells, and the mean surface area of nerve cells/field were significantly reduced, with an increase of dark cells were found in nicotine group compared to control. Immunoreactivity in nicotine group revealed an increase in neuronal α-synuclein, reduction in tyrosine hydroxylase enzyme, an increase in caspase 3 and ultrastructure changes suggestive of neuronal apopto. The blood capillaries were markedly affected. Nicotine induced endothelial and pericytic apoptotic changes, irregular lumena and indistinct endothelial junctional complex. Nicotine administered subcutaneously in a small dose may have a deleterious effect on SNc, mainly involving dopaminergic neurons and blood capillaries. This effect seems to be secondary to an oxidative stress that might be produced by reduced TAC and increased MDA levels.

The Novel Potential Therapeutic Utility of Montelukast in Alleviating Autistic Behavior Induced by Early Postnatal Administration of Thimerosal in Mice.

BACKGROUND AND AIM: Thimerosal (THIM) is a mercury-containing preservative widely used in many biological and medical products including many vaccines. It has been accused of being a possible etiological factor for some neurodevelopmental disorders such as autistic spectrum disorders (ASDs). In our study, the potential therapeutic effect of montelukast, a leukotriene receptor antagonist used to treat seasonal allergies and asthma, on THIM mice model (ASDs model) was examined. METHODOLOGY: Newborn mice were randomly distributed into three groups: (Group 1) Control (Cont.) group received saline injections. (Group 2) THIM-treated (THIM) group received THIM intramuscular (IM) at a dose of 3000 µg Hg/kg on postnatal days 7, 9, 11, and 15. (Group 3) Montelukast-treated (Monte) group received THIM followed by montelukast sodium (10 mg/kg/day) intraperitoneal (IP) for 3 weeks. Mice were evaluated for growth development, social interactions, anxiety, locomotor activity, and cognitive function. Brain histopathology, alpha 7 nicotinic acetylcholine receptors (α7nAChRs), nuclear factor kappa B p65 (NF-κB p65), apoptotic factor (Bax), and brain injury markers were evaluated as well. RESULTS: THIIM significantly impaired social activity and growth development. Montelukast mitigated THIM-induced social deficit probably through α7nAChRs upregulation, NF-κB p65, Bax, and brain injury markers downregulation, thus suppressing THIM-induced neuronal toxicity and inflammation. CONCLUSION: Neonatal exposure to THIM can induce growth retardation and abnormal social interactions similar to those observed in ASDs. Some of these abnormalities could be ameliorated by montelukast via upregulation of α7nAChRs that inhibited NF-κB activation and significant suppression of neuronal injury and the associated apoptosis.

Tramadol administration induced hippocampal cells apoptosis, astrogliosis, and microgliosis in juvenile and adult male mice, histological and immunohistochemical study.

Tramadol is a common analgesic, frequently used for relieving moderate or severe pain and widely used to delay ejaculation. However, repeated large doses have several adverse effects, especially on the brain tissue. So, this study was designed to assess the potentially deleterious effects of chronic administration of tramadol on principal fields of the hippocampus in adult and juvenile male albino mice. Thirty swiss male albino mice were divided equally into three groups: Group Ia (control adult) 3 months old, Group Ib (control juvenile) 3-week postnatal mice, Group II (tramadol treated adult mice) and Group III (tramadol treated juvenile mice). Both treated groups received tramadol tablets dissolved in water in a dose of 40mg/kg for 1 month by gastric tube. Tramadol treated groups showed degenerative changes in dentate gyrus (DG) granule cells, pyramidal neurons of CA1and CA3 fields in the form of electron-dense or rarified cytoplasm, dilated rER and mitochondrial changes. Additionally, immunohistochemical results revealed significantly increased in caspase 3 positive cells in different hippocampal principal fields. Astrogliosis and microgliosis were proved by the increased immunoreactivity of astrocytes to glial fibrillary acidic protein (GFAP) and microglia to CD68. Morphometric findings showed a significant reduction of both surface area of granule and pyramidal cells, and in thickness of DG, CA1, CA3 layers. Moreover, most of these morphological changes were aggravated in the juvenile-treated group. So, it can be concluded that tramadol abuse can induce an altered morphological change on the principal fields of the hippocampus in adult and juvenile mice.

Study of Angiopoietin-2 and vascular endothelial growth factor as markers of diabetic nephropathy onset in Egyptians diabetic patients with non-albuminuric state.

BACKGROUND: Vascular endothelial growth factor (VEGF) and angiopoietin (Ang-2) systems have a central role in vasculogenesis and neoangiogenesis during glomerular development. disruption, their levels are associated with alterations in the glomerular filtration barrier and proteinuria as in diabetic nephropathy. Aim of this study to assess the validity of blood Ang-2 and VEGF as biomarkers for early detection of diabetic nephropathy as well as to study the relation between them and inflammation in diabetic nephropathy patients. SUBJECTS AND METHODS: Cross-sectional study included 180 diabetic nephropathy patients. Patients were classified to non-albuminuric, microalbuminuria and macroalbuminuria patients. Patients with macroalbuminuria complicated to renal impairment and ESRD on top of diabetic nephropathy. Ang-2 and VEGF were measured beside urinary albumin creatinine ratio (UACR). RESULTS: Significant increase of Ang-2 and VEGF levels among patients with normoalbuminuric state compared to control but there is no significant difference of UACR between both groups. Ang-2 and VEGF concentrations were significantly higher in patients with microalbuminuria and macroalbuminuria compared to healthy. Ang-2 and VEGF levels increase with the progression of albuminuria. There were significant positive correlation between CRP and Ang-2 in addition to VEGF. Significant negative correlation between eGFR, Ang-2 and VEGF. CONCLUSION: VEGF and Ang-2 were significantly elevated in diabetic nephropathy patients without albuminuria, their levels steadily increase with the progress of albuminuria, So can use them as markers for diagnosis of diabetic nephropathy onset and progression specially in patients without an increase in albumin excretion.

Rat hippocampal CA3 neuronal injury induced by limb ischemia/reperfusion: A possible restorative effect of alpha lipoic acid.

Limb ischemia reperfusion (I/R) injury is associated with serious local and systemic effects. Reperfusion may augment tissue injury in excess of that produced by ischemia alone. The hippocampus has been reported to be vulnerable to I/R injury. Alpha lipoic acid (ALA) is an endogenous antioxidant with a powerful antioxidative, anti-inflammatory, and antiapoptotic properties. We studied the probable restorative effect of ALA on limb I/R-induced structural damage of rat hippocampus. Forty adult male albino rats were divided equally into four groups: group I (sham); group II (I/R-1 day) has undergone bilateral femoral arteries occlusion (3 h), then reperfusion for 1 day; group III (I/R-7 days) has undergone reperfusion for seven days; group IV (I/R-ALA) has undergone I/R as group III and received an intraperitoneal injection of ALA (100 mg/kg) for 7 days. I/R groups revealed degenerative changes in the pyramidal neuronal perikarya of CA3 field in the form of dark-stained cytoplasm, dilated RER cisternae, mitochondrial alterations, and dense bodies' accumulation. Their dendrites showed disorganized microtubules. Astrogliosis is featured by an increased number and increased immunoreactivity of astrocytes for glial fibrillary acid protein. Morphometric data revealed significant reduction of light neurons, surface area of neurons, and thickness of the CA3 layer. Most blood capillaries exhibited narrow lumen and irregular basal lamina. ALA ameliorated the neuronal damage. Pyramidal neurons revealed preservation of normal structure. Significant increase in the thickness of pyramidal layer in CA3 field and surface area and number of light neurons was observed but astrogliosis persisted. Limb I/R had a deleterious remote effect on the hippocampus aggravated with longer period of reperfusion. This work may encourage the use of ALA in the critical clinical settings with I/R injury.

Protective effect of tetrahydrobiopterin on hepatic and renal damage after acute cadmium exposure in male rats.

Cadmium (Cd) has been recognized as one of the most important environmental and industrial pollutants. This study investigated the impact of acute exposure to Cd on oxidative stress and the inflammatory marker interleukin-6 (IL-6) in the plasma of rats and the histological picture of liver and kidney, as well as to examine the potential protective effect of tetrahydrobiopterin (BH4). METHODS: Rats were divided into control group, Cd group that received a single intraperitoneal (i.p.) dose of 4 mg/kg b.w. of CdCl2 and BH4+ Cd group that received a single dose of BH4 (20 mg/kg, i.p.) and subsequently exposed to a single dose of Cd 24 h after the BH4 treatment. RESULTS: Cd increased the plasma levels of hepatic enzymes (ALT and AST), urea, creatinine, malondialdehyde (MDA), and IL-6 and decreased the superoxide dismutase (SOD) activity. Also, it induced histopathological alterations in the liver with severe degeneration, especially in centrilobular zones. Renal tubular epithelium showed vacuolated cytoplasm and dense nuclei. VEGF expression was mild. Ultrastuctural changes were seen in some renal tubules. The nuclei appeared distorted with electron dense chromatin. Mitochondria with destructed cristae were observed. BH4 pretreatment had protective effects, since it significantly reduced the levels of IL-6 and ameliorated the alteration in oxidative status biomarkers induced by Cd. Improvement of histopathological alterations was observed in Cd-groups. The nuclei were vesicular euchromatic, intact mitochondria and normal appearance of the filtration membrane. Moderate expression of VEGF was noted. CONCLUSION: This study has provided clear evidence for the protective efficacy of BH4 against experimental Cd toxicity.

Nicotine impact on the structure of adult male guinea pig auditory cortex.

BACKGROUND: A growing body of evidence suggests that chronic cigarette smoking causes detrimental effects on brain morphology. AIM OF WORK: To study the structural changes in auditory cortex region (Layer V), under the influence of nicotine. MATERIAL AND METHODS: Three animal groups (10 each) were used; group I (control) and groups IIa and IIb received 3 and 6mg/kg nicotine respectively. The specimens from the auditory cortex were examined using light and electron microscopy and morphometry. RESULTS: Neurons and blood capillaries of the auditory cortex (layer V), were influenced by chronic nicotine treatment in a dose dependent manner. The neurons and their processes revealed disorganization and dissociation of microtubules. The neuronal cells nucleoli characteristically revealed large fibrillar centers detected by silver stain and ultrastructure. The blood capillaries revealed collapse, irregular lumen, thickened basal lamina, abnormal forms of nuclei and organization of microtubules. Neuroglia revealed marked reactivity. Morphometrically, there was a significant decrease in the thickness of the auditory cortex and the number of light neurons and a significant increase in the number of dark neurons in comparison to the control. CONCLUSION: Nicotine affects the integrity of the auditory cortex possibly by reducing metabolic and transcription activities.

Structure of the secretory cells of male and female adult guinea pigs Harderian gland.

The main objective of this study was to investigate the structure of the Harderian gland (HG) in male and female guinea pigs. A total number of sixteen animals of 4 months age were divided according to sex into two groups; eight animals each. Unfixed glands were weighed and their length and width were measured. Specimens from fixed glands were processed and examined using light, transmission electron microscopy and immunohistochemistry for the detection of the presence of chromogranin A (CgA). The gland consisted of a well-developed duct system which included both intra and extra parenchymal ducts and secretory end pieces lined by many types of cells of variable morphological features and modes of secretion. However, the holocrine mode of secretion was rare as mitotic figures were occasionally present. The interstitial cells included fibroblasts and immune cells (mast cells, lymphocyte, plasma cells and macrophages). The secretion produced by the gland included lipid, protein, neutral mucin and CgA which may be a newly identified constituent of biologically potent proteins stored in the cells of the guinea pig HG. Neutral mucin and CgA may function in photoprotection. The gland revealed sexual dimorphism in mast cells and blood capillaries number and chromogranin secretory activity.

Neutrophil CD64 expression in inflammatory autoimmune diseases: its value in distinguishing infection from disease flare.

The purpose of this research was to clarify the significance of neutrophil CD64 expression in discrimination between infection and disease flare in patients with inflammatory autoimmune diseases. The study included 63 subjects, 20 healthy controls and 43 patients with inflammatory autoimmune diseases (24 with rheumatoid arthritis & 19 with systemic lupus erythematosus). The FC gamma receptor I (CD64 expression) on neutrophils was measured using flow cytometry. The intensity of CD64 expression on neutrophils was significantly elevated in patients with infections; 49.0 (13-205), and active autoimmune disease; 36.15 (12-133) compared to healthy controls; 5.35 (2.6-14) or patients with inactive disease; 7.5 (3.3-18). In the infectious disease group, expression of CD64 was significantly higher than in the active inflammatory disease group, while there was no significant difference between the group of patients with inactive inflammatory disease and healthy controls (P > 0.05). The sensitivity of CD64 bearing neutrophil intensity for detection of infection (using a cut off value of ≥43.5) was 94.4% and specificity was 88.9%. Neutrophil CD64 expression has a good sensitivity and specificity in differentiating infection from disease flare in patients with inflammatory autoimmune diseases. This assay could facilitate early and accurate diagnosis and greatly aid timely institution of appropriate treatment.

CD40 ligand, Bcl-2 and apoptosis in B-chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) is a haematopoetic neoplasm caused primarily by defects in apoptosis mechanisms and complicated by progressive marrow failure, immunosupression and increased resistance to chemotherapy. The CD40-CD40 ligand (CD40L) interaction has been shown to significantly increase antigen presentation in normal and malignant B-cells and it is a powerful regulator of cell survival. Bcl-2 expression is common in CLL and is associated with decreased overall survival. Our objective was to asses CD40 ligand (CD154) and Bcl-2 expressions and their correlation with clinical and laboratory features in CLL patients. This study was conducted on 40 subjects, including 10 healthy volunteers as the control group and 30 patients presented with de novo chronic lymphocytic leukemia (CLL), all of them were subjected to thorough history taking, full clinical examinations, routine laboratory investigations and flowcytometric assessment of CD40L and Bcl-2 on lymphocytes. There was a highly significant increase in TLC, absolute lymphocytic count, serum LDH, B2-microglobulin and Bcl-2 expression (P<0.001); there was a significant increase in CD40L expression (P<0.05); whereas there was a highly significant decrease in hemoglobin concentration and platelets count between the study group (P<0.001). There was no significant difference as regard direct Coombs' test between both groups. There was no significant relation between CD154 expression and clinical findings, Rai staging system and other laboratory parameters. CD40L expression is increased with staging of Modified Rai staging system but not reaching the significant level. There was no significant correlation between CD154 expression and some of clinical and laboratory parameters, whereas there was only significantly negative correlation between Bcl-2 expression and both haemoglobin concentration and platelets count (P<0.001). Combination of Bcl-2 antisense oligonucleotide with conventional chemotherapeutic drugs may enhance the cytotoxicity of these drugs and induces apoptosis.