Hamed Benghuzzi

β class II tubulin predominates in normal and tumor breast tissues

BackgroundAntimitotic chemotherapeutic agents target tubulin, the major protein in mitotic spindles. Tubulin isotype composition is thought to be both diagnostic of tumor progression and a determinant of the cellular response to chemotherapy. This implies that there is a difference in isotype composition between normal and tumor tissues.MethodsTo determine whether such a difference occurs in breast tissues, total tubulin was fractionated from lysates of paired normal and tumor breast tissues, and the amounts of β-tubulin classes I + IV, II, and III were measured by competitive enzyme-linked immunosorbent assay (ELISA). Only primary tumor tissues, before chemotherapy, were examined. Her2/neu protein amplification occurs in about 30% of breast tumors and is considered a marker for poor prognosis. To gain insight into whether tubulin isotype levels might be correlated with prognosis, ELISAs were used to quantify Her2/neu protein levels in these tissues.Resultsβ-Tubulin isotype distributions in normal and tumor breast tissues were similar. The most abundant β-tubulin isotypes in these tissues were β-tubulin classes II and I + IV. Her2/neu levels in tumor tissues were 5–30-fold those in normal tissues, although there was no correlation between the Her2/neu biomarker and tubulin isotype levels.ConclusionThese results suggest that tubulin isotype levels, alone or in combination with Her2/neu protein levels, might not be diagnostic of tumorigenesis in breast cancer. However, the presence of a broad distribution of these tubulin isotypes (for example, 40–75% β-tubulin class II) in breast tissue, in conjunction with other factors, might still be relevant to disease progression and cellular response to antimitotic drugs.

Nodular hematopoiesis of the liver diagnosed by fine-needle aspiration cytology

A case of tumor‐like extramedullary hematopoiesis (EMH) of the liver diagnosed by fine‐needle aspiration cytology guided by computer tomography (CT) is reported. The initial clinical diagnosis was metastatic carcinoma from an adrenal gland primary. Five other cases of tumor‐like EMH diagnosed by FNA have been presented in the literature. In two of the cases, the primary clinical diagnosis was metastatic tumor. The most common location for tumor‐like EMH is paravertebral and intrathoracic. Three such cases of paravertebral tumor‐like EMH have been diagnosed by FNA. Nodular EMH can be found rarely in other organs as in the liver. Diagn. Cytopathol. 16:51–54, 1997.

Morphometric Evaluation of Tissue-Implant Reaction Associated With ALCAP and TCP Bioceramics In Vivo

The purpose of this investigation was to correlate the thickness of the fibrous capsule and the various histological components surrounding aluminum-calcium phosphate (ALCAP) and tricalcium phosphate (TCP) bioceramics at the subcutaneous (sc) and intraperitoneal (ip) implantation sites. The rational of conducting this investigation is to further elucidate the mechanisms of tissue-implant interaction. Thirteen Sprague-Dawley adult male albino rats were randomly divided into three groups. Animals in groups I and II (n = 5/group) were implanted at both ip and sc implantation sites with either ALCAP or TCP ceramics, respectively. Animals in group III (n

Characterization of a Femoral Segmental Nonunion Model in Laboratory Rats: Report of a Novel Surgical Technique

The literature is lacking conclusive results regarding the exact mechanism of maximizing the fracture healing stages with minimal traumatic side effects. This observation mandates the development of a novel surgical procedure using small animals as a model to study fracture healing in the presence of osteoinductive agents. Previously, stabilization of osteotomies in small animal models has mainly been accomplished using Kirschner wires, but the rats tremendous ability to heal an osteotomy stabilized by this method has masked the effects of osteoinductive agents. Thus, this study proposes using a modified 20-hole, 1.5-mm stainless-steel plate to stabilize a 5-mm segmental defect. Thirty of 32 adult male rats were fully weight-bearing within 2 days and were followed over a 15-week period. Two animals showed evidence of fixation failure due to technical error, and the animals were humanely sacrificed. At the end of the study, the fractures were stable with significantly less bone formation evident when compared to controls (p <. 001). Therefore, this technique can effectively be used to evaluate compounds that will enhance bone formation and allows for stable fixation of the control with minimal callus formation or bony ingrowth. The goal of this article is to allow other investigators to reproduce this technique as well as outline the advantages and disadvantages of this novel plating technique versus the former Kirschner wire technique for the study of osteoinductive agents using small animals as a model.

Cytological evaluation of the tissue-implant reaction associated with S/C and I/P implantation of ALCAP and HA bioceramics in vivo.

It is well documented that several ceramic materials are highly compatible and non-immunogenic with host tissues. Recent studies have demonstrated the need for further investigation of these devices in vivo to further elucidate the possible mechanisms involved in biocompatibility. The purpose of this investigation was to study the morphological characteristics of the fibrous tissue capsule resulting from the implantation of aluminum calcium phosphate (ALCAP) and hydroxyapetite (HA) bioceramics. Implants of ALCAP and HA were implanted into 10 adult male rats subcutaneously (S/C) and intraperitoneally (I/P). At 90 days post-implantation, the animals were euthanized, and the ceramic devices, the fibrous tissue, and vital organs were harvested. Evaluation of routine stained sections (5 microm, hematoxylin & eosin) of the fibrous tissue capsule surrounding the HA and ALCAP ceramics revealed the following: 1) all the ceramic devices had fibrous connective tissue capsules of slightly varying degrees of thickness at the time of sacrifice, depending on the site of implantation and type of material, and 2) there were statistically significant differences (p < 0.05) in the numbers and types of cellular components with respect to implantation site. The number of macrophages, neutrophils, fibroblasts, degree of vascularity, and thickness of the fibrous tissue matrix was found to be statistically different between the S/C implanted ceramic groups. The number of macrophages, neutrophils, fibroblasts, and collagen content comparing the fibrous tissue surrounding the ALCAP and HA ceramics (I/P), was found to be statisically different.

Suppression of Homocysteine Levels by Vitamin B12 and Folates: Age and Gender Dependency in the Jackson Heart Study

Introduction: To examine factors potentially contributing to premature cardiovascular disease mortality in African Americans (40% versus 20% all other populations), plasma homocysteine, serum vitamin B12 and folate levels were examined for African American participants in the Jackson Heart Study. Methods: Of 5192 African American Jackson Heart Study participants (21–94 years), 5064 (mean age, 55 ± 13 years; 63% female) had homocysteine levels measured via fasting blood samples, with further assessments of participants vitamin B12 (n = 1790) and folate (n = 1788) levels. Regression analyses were used to examine age, gender, vitamin B12 and folate with homocysteine levels. Results: Homocysteine levels, a purported surrogate risk factor for cardiovascular disease, increased with age, were inversely proportional to folate and vitamin B12 levels (P < 0.001) and were higher for men of all ages. Conclusions: The results show that, as with other populations, age, gender, vitamin B12 and folate may predict homocysteine levels for African Americans. Diet may be an important predictive factor as well, given the relationships that were observed between plasma homocysteine and serum B vitamin levels.

The Effects of Long-Term Sustained Delivery of Dihydrotestosterone by Poly(lactic acid) Impregnated and Noncoated Biodegradable Ceramic Devices in Male Rodents

The objectives of this investigation were to evaluate the release of dihydrotestosterone (DHT) from nonimpregnated and poly(lactic acid) (PLA) impregnated ALCAP ceramic reservoirs implanted in male rats, and to study the effects of delivered DHT on the reproductive system of male rats. A total of 120 Sprague-Dawley male albino rats were distributed equally into three groups. Two ALCAP capsules, one nonimpregnated and the other impregnated with PLA, were implanted in each rat in groups I and II. Capsules implanted in group I rats were loaded with 40 mg DHT each. Group II rats were implanted with two empty capsules (sham group), and group III animals served as un implanted controls. Eight rats from each group were euthanized at the end of the one, three, six, nine, and twelve months following the implantation of the ceramics. No significant changes in the weights of vital organs of rats were observed among any of the three different groups. Vas deferens and epididymal fluid were devoid of normal spermatozoa within three months of implanting the steroid-containing ceramics. Testes weights decreased significantly in the rats implanted with ALCAP ceramics containing DHT and the seminiferous tubules became oligospermic after one month and azoospermic after three months. The data collected in this study suggest that: (1) ALCAP ceramic cap sules are capable of delivering DHT for one year at a sustained manner; (2) DHT delivered by ALCAP capsules can be used effectively to regulate sperma togenesis in rats.

Successful antidote of multiple lethal infections using sustained delivery of difluoromethylornithine by means of ceramic drug delivery devices

The objectives of this study were (1) to cure multiple infections of trypanosomiasis in rats by the sustained release of DFMO from biodegradable tricalcium phosphate (TCP) and aluminum-calcium-phosphorous oxide (ALCAP) delivery systems, and (2) to determine if the side effects associated with oral administration of DFMO can be avoided by using TCP and ALCAP capsules. Sixty-eight SD male albino rats (235-270 g) were divided randomly into five groups. Each rat in group I (n = 16) was implanted subcutaneously (s.c.) with four TCP capsules (two large TCP (L-TCP), one PLA-impregnated large TCP (IL-TCP) and one thin TCP capsule (TN-TCP)). Rats in group II (n = 16) were implanted s.c. with four ALCAP ceramics (two large ALCAP (L-ALCAP), one PLA-impregnated large ALCAP (IL-ALCAP) and one thin ALCAP capsule (TN-ALCAP)). Rats in groups III (n = 16), IV (n = 4) and V (n = 16) were left without implants. Rats in group III (n = 16) were given 4% (w/v) DFMO (pH 7) in drinking water at the day of inoculation and continued up to 7 days postinoculation. Rats in group IV (n = 4) served as a nontreated group. Rats in group V (n = 16) served as normal controls. The results showed that all rats implanted with with TCP or ALCAP implants had no intoxications symptoms or side effects such as diarrhea during the treatment period. In contrast, rats given DFMO in drinking water exhibited foul-smelling diarrhea during the treatment period. Microscopic evaluation of blood smears collected from rats receiving DFMO chemotherapy showed an occasional or limited number of stumpy shape (SS) trypanosomes. This study suggests that (1) ceramic drug delivery systems are capable of delivering DFMO in a sustained manner for two months, and were able to cure repeated infections of trypanosomiasis; (2) the use of ceramic implants avoided widely fluctuating, irregular levels of DFMO in the body by keeping sustained levels above minimal effective concentrations; (3) ceramic drug delivery systems provide a pharmacological potentiality for drugs such as DFMO which have been withheld from the market because of severe side effects when administered using conventional methods of drug administration; and (4) DFMO-filled ceramic devices can be implanted subcutaneously in animals that face a threat of lethal protozoal infections in highly infested areas of the world.

A preliminary report on the effects of sustained administration of corticosteroid on traumatized disc using the adult male rat model.

Study Design A novel degenerative disc disease model and sustained delivery method for corticosteroid in male Sprague-Dawley albino rats. Objectives To develop a model of degenerative disc disease and to determine the effect of continuous sustained release of corticosteroid on the process of degeneration within the traumatized disc. Summary of Background Data The current modalities of treating symptomatic degenerative disc disease are either conservative or surgical. However, there is no cure for the degenerative process and prevention, therefore, is the ideal treatment. An understanding of the mechanisms involved in disc degeneration is crucial to develop new methods for prevention and treatment, including appropriate delivery systems and dosages of repair factors. Methods The L5-L6 intervertebral disc was pierced with a 23-gauge needle in 18 rats. The animals received either sham or corticosterone-charged tricalcium phosphate ceramic capsules. The rats were euthanized at 4 weeks. Chondrocytes in the transition zone areas were counted and compared statistically. Results The surgical technique induced degeneration of the nucleus without evidence of inflammation at adjacent levels when compared with nontraumatized controls. The number of chondrocytes per area was significantly less in the sham group than in the control group. Corticosteroid treatment showed chondrocyte numbers similar to control in 4 of 5 different views of the disc. The anterior region of the disc had 50% less chondrocytes per area than the control; however, the chondrocyte numbers were 50% greater than in the same site from discs of sham animals. Conlcusions The results show the development of a degenerative disc animal model that can be used to test the effects of growth enhancing factors in disc repair. Administration of continuous sustained release of corticosterone can slow the process of degeneration within the traumatized disc in the rat model.

Altered neuro-inflammatory gene expression in hippocampus in major depressive disorder

&NA; Major Depressive Disorder (MDD) is a common psychiatric disorder for which available medications are often not effective. The high prevalence of MDD and modest response to existing therapies compels efforts to better understand and treat the disorder. Decreased hippocampal volume with increasing duration of depression suggests altered gene expression or even a decrease in neurogenesis. Tissue punches from the dentate gyrus were collected postmortem from 23 subjects with MDD and 23 psychiatrically‐normal control subjects. Total RNA was isolated and whole transcriptome paired‐end RNA‐sequencing was performed using an Illumina NextSeq 500. For each sample, raw RNA‐seq reads were aligned to the Ensembl GRCh38 human reference genome. Analysis revealed 30 genes differentially expressed in MDD compared to controls (FDR < 0.05). Down‐regulated genes included several with inflammatory function (ISG15, IFI44L, IFI6, NR4A1/Nur‐77) and GABBR1 while up‐regulated genes included several with cytokine function (CCL2/MCP‐1), inhibitors of angiogenesis (ADM, ADAMTS9), and the KANSL1 gene, a histone acetyltransferase. Similar analyses of specific subsets of MDD subjects (suicide vs. non‐suicide, single vs. multiple episodes) yielded similar, though not identical, results. Enrichment analysis identified an over‐representation of inflammatory and neurogenesis‐related (ERK/MAPK) signaling pathways significantly altered in the hippocampal dentate gyrus in MDD. Together, these data implicate neuro‐inflammation as playing a crucial role in MDD. These findings support continued efforts to identify adjunctive approaches towards the treatment of MDD with drugs including anti‐inflammatory and neuroprotective properties. HighlightsIn MDD, 30 genes are differentially expressed in dentate gyrus compared to controls.MDD is associated with changes in genes with neuro‐inflammatory and angiogenesis functions.Inflammatory and neurogenesis‐related signaling pathways are altered in MDD.Anti‐inflammatory treatments may be an adjunctive therapeutic approach to MDD.