Immuno Expression Research Articles

Putative role of 6-chogaol against tramadol-induced hepatotoxicity in albino rats via anti-inflammatory, antifibrotic, and antiapoptotic effects

Tramadol is a commonly used drug to relieve pain and avoid premature ejaculation in males with hepatotoxic effects, and 6-chogaol has potent anti-inflammatory and hepatoprotective properties.The work impetus is probing the hepatoprotective mechanisms of 6-chogaol against tramadol hepatoxicity. Twenty adult male rats were enrolled to obtain four equal groups [control group (G1), 6-chogaol group (G2), tramadol group (G3), and 6-chogaol+tramadol group (G4)]. Liver specimens were excised and processed to evaluate hepatocyte injury through histopathological (HP), immunohistochemical (IHC), flow cytometry, and biochemical investigations. The HP study exhibited hepatic injury in G3 hepatocytes (inflammatory cell infiltration, hepatic fibrosis, and disturbed liver structure). The IHC study showed a significant rise in caspase-3 and reduced PCNA immuno-expression (IE). Likewise, the flow cytometry and biochemical experiments exhibited a substantial elevation of apoptotic hepatocytes and the serum levels of IL-1β, IL-6, TNF-α, ALP, ALT, and AST in G3. In contrast, G4 rats significantly improved in all HP, IHC, flow cytometry, and biochemical parameters. Collectively, tramadol intake exerted harmful toxic effects on hepatocytes, whereas 6-Shogaol hampered these changes and served as a natural hepatoprotective agent. Therefore, we advise concurrent intake of 6-Shogaol supplement with tramadol to preserve the integrity of hepatic tissues.

Immunohistochemical Expression of CDX2 as Early Biomarkers for Gastric Intestinal Metaplasia among Sudanese Patients with Chronic Gastritis

Background: Chronic gastric infection with Helicobacter pylori can cause intestinal metaplasia, a pathologic change that is frequently a precursor of gastric cancer; CDX2 has an essential role in the development and maintenance of intestinal differentiation in the gut and ectopic sites such as intestinal metaplasia of the stomach. Objective: This study aimed to investigate caudal-type home box (CDX2) Immuno expression as an early indicator for gastric intestinal metaplasia among Sudanese patients with chronic gastritis. Materials and methods: This is a descriptive retrospective cross-sectional study conducted at Atbara Medical Complex and El-MakNimr University Hospital at River Nile State in the period from 2019 to 2022. 140 archival paraffin tissue blocks were collected from patients diagnosed with chronic gastritis, immunohistochemistry was used to detect (CDX2 expression as an early indicator for gastric intestinal metaplasia. Results: From 140 samples, 80 were females (57.1 %) and 60 were males (42.9%), the patient's ages ranged from (12 to 97) with a mean age of (53.27) years, and age was categorized into three groups, less than 40 (27) (19.3%), 40- 60 (64) (45.7%), more than 60 (49) (35.0%). The prevalence of gastric intestinal metaplasia was (17.9%, while no statistically significant correlation of gastric intestinal metaplasia with gender and age as the P.value was (0.308 and 0.256) respectively. Conclusion: The development of gastric intestinal metaplasia as a premalignant condition of gastric carcinoma is important to detect this change at an early point to prevent and control gastric cancer.

Ki-67 IMMUNOEXPRESSION IN ADENOCARCINOMA OF PROSTATE AND ITS CORRELATION WITH GLEASON GRADE

Background: Prostate cancer represents the second most frequently diagnosed malignancy among the male and fth leading cause of death. Tumour grade is one of the most important prognostic factors of prostate cancer. There are many markers studied in prostatic cancers, but the most extensively studied one is Ki-67, the proliferative marker. Prostatic cancers with Ki-67 over expression are generally aggressive. Aim And Objectives: - Aim of this study is evaluate ki67 immunoexpression in adenocarcinoma of prostate and to correlate with histopathological grade (Gleason Grade). This is single centre cross sectional study. The patie Material Methods:- nt who visited department of urology and renal transplant in Gauhati medical college and hospital and whose TRUS guided biopsy were received in the pathology department and diagnosed as adenocarcinoma were included. A total 50 cases were analysed. Tumour grade was determined according to Gleason's grading system and Ki-67 expressions were determined by IHC staining. The obtained results were analysed and evaluated using Chi square test (IBM SPSS version 21). Result And Interpretation:- A statically signicant correlation found between Gleason Grade and Ki-67 immuno expression with a p value of <0.001. In this study a statistically signicant correlation was found between Conclusion: - Gleason Grade and ki-67 labelling index. Hence Ki-67 immunoexpression evaluation and its result can be incorporated with histopathology which may inuence clinical decision making as prognosis of prostate cancer.

Immunohistochemical Expression of NKX3.1 in Prostatic Adenocarcinoma Correlates with Tumors Grade among Sudanese

Background: The second most frequent cancer in males and the fourth most common cancer overall for both sexes is prostate cancer. On chromosome 8p, there is a prostatic tumor suppressor gene called NKX3.1. Despite the fact that the majority of primary prostatic adenocarcinomas exhibit positive NKX3.1 protein staining. Objective: The purpose of this study was to evaluate the immunohistochemical expression of NKX3.1in prostate adenocarcinoma and correlate it with tumor grades among Sudanese patients. Materials and Methods: This study is a retrospective descriptive cross- sectional study, was conducted in Khartoum state, (Radiation& Istopes Center- Khartoum (RICK)). Tissue sections were stained by immunohistochemical staining against NKX3.1 (indirect dextral polymers (Dako- EnVision TM Flex kit)) was carried out on forty (40) archival formalin fixed paraffin embedded tissue blocks from patients diagnosed as prostate carcinoma. Data were collected from hospital records and the immunohistochemical results and then analyzed using SPSS 25.0 frequency, and Chi-Square and mean were calculated. Results: The study found that the most frequent age group is between (71 – 80) years old, twenty (20) cases have prostate adenocarcinoma grade III, twelve (12) cases were grade II, and eight (8) cases were grade I. Any nuclear NKX3.1 staining was regarded as positive. Thirty-five (35%) have NKX3 positive immunostain while (65%) have a negative result. There is a statistically significant correlation between the NKX3.1 expressions and tumor grade as the P. value was (0.000), while in a statistically significant correlation between the NKX3 expressions and patients' age as the P. value was (0.957). Conclusions: NKX3 immuno expression is strongly associated with higher tumor grade and may prove the role of this protein in the progression of prostate cancer.

Effect of Zinc in Submandibular Salivary Gland Branching Morphogenesis

IntroductionNerves from the parasympathetic branch are critical for the function of the glandular tissue including salivary gland. Parasympathetic innervation influences organ regeneration: tissue regrowth impaired after parasympathectomy in the salivary gland. As it is difficult to treat the damaged nerves, there is a need to develop new materials and treatment methods to treat the nerves. Zinc ions are reported to be important metal ions in the physiological functions of the nervous system and involved in neural cell proliferation and differentiation. Here, we aimed to study the influence and neuroprotective role of the extracellular zinc during submandibular salivary gland (SMG) development using ex vivo tissue culture system. Also we prepared zinc ion‐containing agarose hydrogel beads (Zn+Aga) for sustained release of zinc ion.Materials and Methods300~500µm size beads prepared. Different concentration of zinc were incorporated in the beads. LY294002 an antagonist of PI3Kinase pathway also incorporate as needed. Next, 0.3 wt%~4 wt% alginate hydrogel sheets prepared for ex‐vivo organ culture. SMG were isolated from the pregnant ICR mouse at day 13 (E13) and culture on hydrogel sheet with or without Zn ion and LY294002 containing beads. Finally, the data analyzed histomorphometrically.ResultsWe successfully prepare the Zn+Aga and observed the sustain release of Zn+ upto 48 hours. Histological evaluation showed that zinc ion promoted the growth of submandibular tissue. The immuno expression of AKT was promoted by the administration of zinc ion. AKT decreased by using ly294002, which is an inhibitor of PI3kinase, and it recovered after administration of zinc. This indicates that zinc administration is involved in the AKT‐pathway.Discussion & ConclusionCollectively, these data clearly illustrate that the Zn is essential element during salivary gland organogenesis through activation of the PI3Kinase‐AKT pathway.

The protective effect of Spirulina versus vitamin E on Electromagnetic waves induced thyroid gland damage in the adult albino rat: a histological and immunohistochemical study

Introduction: widespread use of several electrical and telecommunication applications that produce EMFs is increasing rapidly. One of the most important organs which affected by numerous types of EMWs is the thyroid gland.Aim of the study: This study targets to explore effects of EMW exposure on the Thyroid and the protective influence of spirulina platensis and vit E . Material and methods: fifty healthy adult male albino rats were separated into five equal groups. Control group, EMW-exposed at which, rats were subjected to the effect of EMF 4 hours daily for a period of one month . EMW exposed- Spirulina treated group at which rats received Spirulina at a dosage of 300 mg/kg b.wt. just previous to EMW exposure daily for one month. EMW exposed - Vit E treated group at which , rats were subjected to EMF like that of group II and given Vit E orally at a dosage of 200 mg/kg. b.wt and a recovery group at which rats kept for one month after one month exposure to low frequency EMF. then The thyroid tissues were examined for histopathological and immunohistochemical changes. Results: group subjected to EMW showed disorganized & degenerated thyroid follicles .Numerous collagen fibers were present within the CT septa in the masson stained thyroid sections . Amount of PAS positive central colloid showed obvious decrease, with Strong KI-67 immunoreactivity and TGFᵦ1 immuno expression. Spirulina platensis and vit E reduces the effect of EMW on thyroid , but vit E revealed obvious increase in the PAS positive central colloid , significant decrease in collagen fibers, KI-67 immunoreactivity and TGFᵦ1 immuno expression compared with that in spirulina group.Conclusion: use of spirulina platensis during period of EMW exposure considered to have a protecting influence. The utilization of vit E has a more protection than spirulina

Potential Chemopreventive Role of Boldine Against Hepatocellular Carcinoma via Modulation of Cell Cycle Proteins in Rat Model.

To evaluate the chemopreventive potential of boldine against diethylnitrosamine (DEN) induced hepatocellular carcinoma (HCC) in Wistar albino rats. Boldine is an alkaloid isolated from Peumus boldus. The primary active constituents of boldine exhibited several potential medicinal properties. The present study was evaluated to explore the chemopreventive agent of boldine on anti-proliferative efficacy against diethylnitrosamine (DEN) induced hepatocellular carcinoma (HCC) in Wistar albino rats. The effect of boldine on cellular proliferative markers, i.e., PCNA and Ki67on hepatocellular carcinoma rats was determined by immuno expression study. Liver marker enzymes, tumor biomarker, oxidative stress markers, antioxidant status, and xenobiotic phase I and II enzymes in HCC rats were analyzed. Moreover, cell cycle proteins, i.e., p21Cip1/Kip1and p27 Cip1/Kip1, Cyclin D1, CDK 4, Cyclin E1, and CDK 2 were investigated using immuno expression analysis. Treatment of boldine protected the liver against reactive oxygen species such as hydrogen peroxide, superoxide, protein carbonyl, and lipid peroxide during hepatocarcinogenesis by boosted antioxidants-superoxide dismutase (SOD), catalase (CAT). Boldine caused a substantial enhanced detoxification process by moderating phase I and II xenobiotic-metabolizing enzymes. Besides, the study found that boldine significantly inhibited the cellular proliferative markers like PCNA and Ki67 and regulated the specific cell cycle-associated proteins by up-regulated expression of p21Cip1/Kip1and p27 Cip1/Kip1 and down-regulated expression of Cyclin D1, CDK 4, Cyclin E1, and CDK 2. Our data manifests the anti-proliferative effect of boldine, which negatively modulates cellular proliferation and regulates cell cycle by protecting the cell from reactive oxygen species (ROS), suggesting that boldine establishes it as a chemopreventive agent in diethylnitrosamine-induced hepatocarcinogenesis in rats.

Expression of Carbonic Anhydrase-IX and Vascular Endothelial Growth Factor in Renal Cell Carcinoma and their Prognostic Significance

Introduction: Renal Cell Carcinoma (RCC) is the most common adult renal malignancy. Histopathologically, clear cell RCC accounts for 65-70% of all RCCs. Carbonic Anhydrase-IX (CA-IX) is a transmembrane protein and takes a role in cancer development and progression. A 75-100% clear cell RCCs show CA-IX expression. But this varies with grade and stage of tumour. Vascular Endothelial Growth Factor (VEGF) is responsible for tumour angiogenesis and expressed variedly in RCCs. Both VEGF and CA-IX expression is mediated by Hypoxia Inducible Factor-1α (HIF-1α). Aim: The present study aimed to evaluate the expression of CA-IX and VEGF in respect to different grades and stages of RCC and assessing their prognostic significance. Materials and Methods: This was a cross-sectional, observational study done on 45 histopathogically diagnosed cases of RCC. It was performed in the Department of Pathology, Nil Ratan Sircar Medical College, Kolkata, West Banglore, India over a period of two years (February 2018 to January 2020). Expression of VEGF and CA-IX were studied by immunohistochemistry. Results were analysed in Statistical Package for Social Sciences (SPSS) software (version 16.0) using Pearson Chi-square test. A p-value of <0.05 was regarded as significant. Results: Out of 45 cases of RCC, 34 tumours (32 clear cell carcinoma and two multilocular cystic renal neoplasm of low malignant potential) were evaluated for CA-IX immuno expression. About 25 cases showed CA-IX positivity which inversely associated with grade and stage of RCC (p-value <0.05). The CA-IX had a diagnostic value in detecting clear cell RCC with sensitivity 73.53%, specificity 100% and accuracy 80%. With 28 (62.2%) cases of RCC showed VEGF positivity among which nine were VEGF 1+ and 19 cases VEGF 2+. The VEGF expression showed a positive association with the grade and Tumour-Node-Metastasis (TNM) stage of tumour (p-value <0.05). Finally, authors found a statistically significant inverse association between CA-IX and VEGF expression in RCCs with clear cell morphology including clear cell RCC and multilocular cystic renal neoplasm of low malignant potential (p-value=0.001). Conclusion: High grade RCCs show low expression of CA-IX and strong positivity with VEGF. Both these markers have a prognostic significance. From the therapeutic point of view, VEGF positive tumours, especially inoperable and metastatic cases, may be benefited by anti-VEGF therapy whereas CA-IX positive tumours respond well by treatment with Interleukin-2.

A study on expression of p16INK4a by immunohistochemistry in cervical dysplasia and Invasive carcinoma of cervix

Introduction: Cervical cancer is the most common cancer in developing country. Early diagnosis of intraepithelial cervical neoplasia and malignancy is important. Infection with Human Papilloma Virus (HPV) results in precancerous lesions and invasive cervical cancer. Interaction of HPV oncogenic proteins with cellular regulatory proteins leads to upregulation of p16INK4a, a cyclin dependent kinase inhibitor. Therefore, p16INK4a overexpression is a surrogate biomarker of HPV infection making it useful in evaluating HPV associated premalignant and malignant lesions of gynecological tract. In our study we have used p16INK4a IHC staining to find out its correlation with dysplasia grading and cervical carcinoma. Materials and Methods: This retrospective descriptive case series for five years duration (from Jan 2014 to Dec2018) was conducted in the Department of Pathology, IGMC & RI, Puducherry, India. A total of 43 cervical biopsies and hysterectomies were collected out of which 17 cases were of carcinoma cervix and 26 cases were of cervical intraepithelial neoplasia ( CIN). All biopsies were subjected to p16INK4a staining. Results: The Immuno expression of p16INK4a of CIN I was 13.3%, CIN II was 20%, CIN III was 50% and Carcinoma cervix was 100%. In normal (non dysplasia) cervical epithelium it showed, 100% negative association. Conclusion: IHC expression of p16INK4a in cervical biopsy is directly related to degree of histological dysplasia and malignancy, so it may be used as a adjunct with routine histopathological examination in identifying cervical dysplasia and also has prognostic value in the management of cervical lesions. Keywords: Dysplasia, Cervical cancer, p16INK4a CIN

Immuno expression pattern of proliferating cell nuclear antigen (PCNA) in normal and atretic follicles of buffalo ovary

The present study was aimed to determine the expression pattern of PCNA in normal and atretic follicles of buffalo ovary. PCNA was localized specifically in the nucleus of the different types of cells in the ovary of buffalo. It was localized in the follicular cells of primordial and primary follicles, granulosa cells in secondary follicles and in all the compartments of a healthy tertiary follicle. The atretic follicles revealed weak PCNA expression. PCNA expression varied in different degrees of atresia and during different seasons.

The Possible Protective Role of Sesame Oil on Skeletal Muscle Regeneration in Induced Red-Bull Injury: Experimental Study

Background: Energy drinks (Eds), including Red Bull, were introduced to United States markets in 1997. Consumption hasincreased worldwide mainly consumed by athletes and teenagers as they are seeking for increase their energy level. Seasme oil(SO) has been known to have anti-inflammatory and antioxidant properties.Aim: The current study was designed to elucidate study the possible protective role of SO on skeletal muscle regeneration inRed Bull experimentally induced injury.Materials and Methods: Twenty adult albino rats were divided into four groups, control, sham control, RB and RB+SO.Skeletal muscle sections were subjected to histological, morphometric, biochemical and statistical studies.Results: RB induced various histological changes in skeletal muscle in the form of congestion, atypical fibre, multiple fibroblastand dark nuclei. RB also caused a sig increase in area % collagen, area % of caspase3 immunoexpression (IE), count of alphasmooth muscle actin IE(no I SMA) and count of +ve CD34 IE were recorded. Sig increase in the activity of interleukin-6(IL-6)and PI3K and AKT proteins. Concomitant administration with SO improved sig the previously mentioned changes.Conclusion: It can be concluded that SO enhanced skeletal muscle regeneration in RB induced muscle injury expressed as adefinite ameliorating effect to the induced inflammatory and degenerative changes.

Ameliorative Role of Melatonin on CA1 of Hippocampus and Frontal Cortex in Induced Social Isolation in Adult Male Albino Rat

Abstract Background: Social Isolation (SI) which means absence or insufficient contact with others is a key factor in psychiatric disorders development, which might be necessary for effective psychological treatment as social rehabilitation. Social expe-rience during the infantile period affects brain function and behaviour in rodents, dogs and human. Aim of Study: The current study was designed to elucidate the neuroprotective role of MT on experimentally-induced SI on CA1of rat hippocampus and frontal cortex. Material and Methods: Twenty adult albino rats were divided into four groups, control, sham control, SI and SI + MT. The Body Weight (BW), Brain Weight (BrW) were determined. Brain sections were subjected to histological, morphometric, biochemical and statistical studies. Results: SI induced various histological changes in both hippocampus and frontal cortex in the form of degeneration and absent neurons. SI also caused a significant (sig) increase in area % of caspase 3 and TNF Immunoexpression (IE) were recorded. Sig decrease in the activity of Total Antioxidant Capacity (TAC), Brain Derived Neurotropic Factor (BDNF), Superoxide Dismutase (SOD) activity, Acetyle Choline Este-rase (ACHE), dopamine level as well ARC gene, and a sig increase in Malondialdehyde (MDA) levels, FOS gene, sero-tonin and AB-42. Concomitant administration with MT im-proved sig the previously mentioned changes. Conclusion: SI might lead to various neurological degen-erations as indicated by the histological, morphometric, biochemical changes. These changes were ameliorated by concomitant administration with MT.

Prognostic value of stem cell markers in glioblastoma

Background/Context: Glioblastoma (GB) is the most common primary brain tumour in adults and it is associated with a high mortality rate. According to the stem cell theory, the growth, relapse and treatment response of GB is determined by the stem cell subpopulation present in the tumour.Objective: Our aim is to study the prognostic value of stem cell markers (CD44, Nestin, Olig2 and SOX2) in a series of homogeneously treated GBs.Material and methods: We study 280 GBs treated with STUPP acheme with a histologican review of the cases and TMA with a máximum of 4 spots for each case. Each slide was immunohistochemically stained and Reading. We compared the immunohistochemical results with survival tme.Results: Only SOX2 immunoexpression (IE) excedding 10% of the tumour cells was found to be related to good survival (p= 0.037) in univariate analysis. However, amultivariate analysis indicate the age, surgery and MGMT promotes methylation but no SOX2 IE are prognostic factors.Conclusions: We conclude the immunohistochemical studies of stem cell markers in GB are not useful for predicting prognosis in daily practice.

Possible Role of Selenium Nano-particles on Gentamicin-Induced Toxicity in Rat Testis: Morphological and Morphometric study

Background: Gentamicin (GM) is abroad spectrum antibiotic that used worldwide. Exact mechanism of its testicular toxicity is still unknown. The present study aimed at studying the possible protective role of selenium nano-particles (SeNPs) on histological, biochemical, and morphometric changes in rat testis.Aim of Study: The current study was designed to elucidate the possible role of SeNPs on GM induced toxicity in rat testis.Materials and Methods: Twenty adult albino rats were divided into four groups, control, sham control, GM group, GM+ SeNPs group. The body weight (BW), testis weight, epididymis weight , seminal vesicle weight, prostate weight and serum testosterone level were determined. Testis sections were subjected to histological, morphometric, biochemical and statistical studiesResults: GM induced significant (sig) reduction in weight of testes, accessory sex organs, sperm count, motility and testosterone level. The drug also caused sig decrease in the activity of testicular catalase (CAT) and reduced glutathione (GSH) content, and sig increase in malondialdehyde (MDA) and H2O2 levels. Sig increase in basement membrane (BM) thickness, area % of caspase3 immunoexpression (IE) and in Ki67 count were recorded. Injection of SeNPs improved sig the previously mentioned changes.Conclusion: GM induced morphological, morphometric and biochemical changes on reproductive organs particularly testis of adult male rats. SeNPs administration with GM ameliorated these adverse effects that might be due to its antioxidant activity.

Nemaline myopathy and secondary mitochondrial dysfunction of complex I

Introduction: Nemaline myopathy (NM) is a heterogeneous muscular congenital myopathy that is slowly progressive or non-progressive. Case report: Our patient was born at 41+6 weeks of gestation, following an uneventful pregnancy .On day four, she presented with weight loss of >10% and hypotonia. The results of the investigations were all normal: investigations included creatine kinase, plasma amino-acids, ammonia level, acylcarnitine profile, lactate and blood gases. Brain magnetic resonance imaging at 3 months of age was normal. Genetic screening for Spinal muscular atrophy (SMA1), myotonic dystrophy (DM1), congenital myasthenic syndrome and Prader-Willi syndrome were all negative. Her echocardiography was normal. Electromyography and nerve conduction studies of the biceps and rectus femoris showed normal sensory responses and there was polyphasic motor units of low amplitude and short duration in the biceps and rectus femoris. The findings suggested a primary muscular disorder of unknown origin. Her skeletal muscle biopsy from right quadriceps which was done at the age of one year showed “thread like” inclusions seen both on light and electron microscopy confirming the diagnosis of NM. The histology also showed normal mitochondria in placement, number and morphologic structure. In addition it showed a moderately low enzyme level of complex I. Mitochondrial whole genome sequencing using the patients skeletal muscle-derived DNA sample detected a known pathogenic mutation, m4298 G>A, at a very low level of heteroplasmy (approximately 10%) in the MTTI (mt-tRNA) gene. The subsequent genetic testing for the most frequent genetic variants associated with nemaline myopathy showed heterozygosity for ACTA1 c.760A > C p.(Asn 254 His). The ACTA1 sequence variant has not been previously reported in the literature. Further assessment of her skeletal muscle biopsy using a quadruple immune-histochemical assay indicated normal levels of both NDUFB8 and COX1 immuno expression. Conclusion: We are describing a severe neonatal onset phenotype of genetically confirmed congenital NM.

Analysis of GPER1, CXCR1 and HER-2 Immuno Expression in Follicular Thyroid Lesions and their Possible Role as Diagnostic Markers

Analysis of GPER1, CXCR1 and HER-2 Immuno Expression in Follicular Thyroid Lesions and their Possible Role as Diagnostic Markers

Resistance to Trastuzumab in Clinical Her2 Breast Cancer: Possible Relation to Molecular Subtype and Cancer Stem Cells

Molecular breast cancer (BC) subtypes and cancer stem cells (CSCs) can determine sensitivity to trastuzumab. A better predictivevalue of clinical human epidermal growth factor receptor2 (cHER2) BC can be provided, by incorporating CSCs heterogeneityinto clinical decisions. The study aimed at investigating involvement of transition of CSCs biology and the molecular subtype indevelopment of trastuzumab resistance in HER2 BC, recommending the use of trastuzumab combinations/modifications able tosuppress CSCs and overcome trastuzumab resistance. The study was conducted on 40 BC patients age range 27-63 years, 20served as control group and 20 as metastatic (predicted resistance) group. Clinicopathological analysis included HER2, ER andPR status, chemotherapy regimen and number of herceptin cycles. Histological analysis included, molecular subtype markers(HER2 and cytokeratin (CK) 5/6), CSCs marker CD44 and morphometric studies. Quantitative polymerase chain reaction (qPCR)was performed. In the metastatic group significant decrease in the area% of HER2 positive (+ve) immunoexpression (IE),significant increase in the area% of CK5/6 IE and CD44 +ve cells was detected. qPCR values of HER2 gene confirmed asignificant decrease. It was concluded that resistance to trastuzumab in the metastatic group of HER2 BC is related to basal likesubtype and overexpression of CD44 +ve CSCs. This nictitates IHC assessment of molecular subtype and type of CSCs to allowthe choice of most appropriate treatment regimen giving best expected prognosis.

HISTOLOGICAL AND IMMUNOHISTOCHEMICAL EVALUATION OF THE POSSIBLE EFFECTS OF DIFFERENT DOSES OF CAFFEINE ON SUBMANDIBULAR SALIVARY GLAND OF ADULT MALE ALBINO RATS USING (ANNEXIN A5) MARKER

Background :Caffeine is one of the most widely consumed substances in the world. The main sources of caffeine in society are coffee or tea The cytotoxicity of caffeine may be due to its ability to trigger apoptosis Material and methods :A total number of 30 adult male albino rats (150-200 g) were used and were divided randomly into five equal groups (n=6 ) control group: 6 rats were I.P injected with sterile: Group II & IIa :12 rats were I. P. injected with single daily dose (2.5 mg/100g body weight/day) for three months this dose is equivalent to minimum human dose. Group IIa: 6 rats stopped treatment and left for one month after the period of treatment.Group III & IIIa: 12 rats were I.P. Injected with single daily dose (10 mg/100g body weight/day) for three months this dose is equivalent to maximum human dose. Group IIIa: 6 rats stopped treatment and left for one month after the period of treatment Samples from the submandibular salivary glands were fixed in 10% buffered neutral formalin and prepared routinely for paraffin sectioning and staining for histopathological changes and immuno- histochemical investigations of apoptotic cell antigen (Annexin A5). Results : Histopathological examination of caffeine treated rats revealed marked changes in glandular architecture, the acini became shrunkn with loss of their rnormal circular arrangement and their cytoplasm was vacuolated. Some serous cells degenerated and replaced by eosinophlic material. The nuclear changes appeared in form of (pleomorphism, hypertrophy, hyperchromatism and pyknosis), numerous normal and abnormal mitotic Figures were detected. Intercalated and excretory ducts were dilated. , blood vessels were dilated and congested. The immuno expression of Annexin A5 in the cytoplasmic basement membrane of the ductal cells was intense and highly significant. Changes were more aggressive in (group III). Recovery of some histological changes were observed especially in group received lower dose of caffeine . Conclusion: Excessive use of Caffeine markedly affect the histological structure of submandibular salivary gland. These changes could be reversible after caffeine stoppage if the doses is small while the histopathological changes of the gland is irreversible if the doses is large .

Immuno Expression of PAKs in Actinic Cheilitis and in Lip Squamous Cell Carcinoma

Lip squamous cell carcinoma (LSCC) may develop from a potentially malignant disorder known as Actinic cheilitis (AC). The p21-activated kinases (PAKs) are possible targets for cancer therapeutics. The LSSC and AC samples were analyzed by immunohistochemistry in order to detects endogenous levels PAKs group I (PAK1 and PAK2) and group II (PAK4, PAK5 and PAK6) only when they were in phosphorylated form (active). Keratinocytes of the basals stratum showed intense staining for phosphoPAKs 1/2, while the most superficial cells of the stratum spinosum and the surface layer showed moderate immunostaining for PAKs 4/5/6 phosphorylated in the epithelium adjacent to injured area. AC did not immunoexpress actives PAKs 1/2 and PAKs 4/5/6. LSSC did not immunoexpress phosphoPAKs 1/2, but some degenerating cells and cells in the necrotic area showed intense staining. Our results suggest that PAKs 1/2 not participate in the regulation of AC and LSCC pathogenesis, while PAKs 4/5/6 are involved in the regulation of cell death in LSCC.

Impact of Chamomile on Submandibular Salivary Gland of 5-Fluorouracil Treated Rabbits (Histological and Immunohistochemical Study)

Background and objectives: Chamomile has been used for centuries as an anti-inflammatory, antioxidant, and mild astringent and healing medicine. The purpose of this study was to investigate the potential anticytotoxic effect of chamomile extract on the submandibular glands of 5-FU treated female rabbits. Materials and methods: Forty female white rabbits weighing between 1.0 kg and 1.5 kg were divided randomly into two groups. Control groups (intraperitoneally injected by physiological saline) consist of distilled water treated group and chamomile extract treated group (100 mg/kg), 10 animals each, and the treatment continue for 16 days. Study groups (intraperitoneally injected by 5-FU) consist also of distilled water treated group and chamomile extract treated group, 10 animals each, and the treatment also continue for 16 days. They were gavaged by distilled water or by chamomile extract in a similar way like control groups, but intraperitoneal injections of the 5-FU at a dose of (4 mg/kg) administered intraperitoneally once daily for 5 successive days. The right and left submandibular salivary glands of all the animals were dissected out and prepared for histological and immunohistochemical examinations. \ Results: 5-FU cause signs of acini degeneration and large dilated blood vessels engorged with red blood cells were also seen. A statistically no significant differences were found between saline/distilled water and the other three groups regarding Ki-67 immuno expression (p>0.05). Statistical analysis also showed no significant differences present between saline/distilled water and saline/chamomile group in term of rate of apoptosis (p>0.05), but this difference were significant (p<0.05) between group 5-FU/distilled water and 5-FU/chamomile group, saline/ distilled water group and 5-FU/distilled water or 5-FU/chamomile group (p<0.05), and saline/chamomile and 5-FU/ distilled water or 5-FU/chamomile group. Conclusion: Chamomile extracts at a concentration of 100 mg/kg/day orally gavaged for 16 days cause some microscopically changes to rabbit’s submandibular salivary gland. Cytotoxicity of chamomile extract increased if it’s taken with 5-FU by increasing the expression of cleaved caspase-3 immunostaining.