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R Guzman

Publications and source records attributed to R Guzman.

At least 55 records · Page 3Linked to original sources

Comparison of mesencephalic free-floating tissue culture grafts and cell suspension grafts in the 6-hydroxydopamine-lesioned rat.

Ventral mesencephalon (VM) of fetal rat and human origin grown as free-floating roller-tube (FFRT) cultures can survive subsequent grafting to the adult rat striatum. To further explore the functional efficacy of such grafts, embryonic day 13 ventral mesencephalic tissue was grafted either after 7 days in culture or directly as dissociated cell suspensions, and compared with regard to neuronal survival and ability to normalize rotational behavior in adult rats with unilateral 6-hydroxydopamine (6-OHDA) lesions. Other lesioned rats received injections of cell-free medium and served as controls. The amphetamine-induced rotational behavior of all 6-OHDA-lesioned animals was monitored at various time points from 18 days before transplantation and up to 80 days after transplantation. Tyrosine hydroxylase (TH) immunostaining of the histologically processed brains served to assess the long-term survival of grafted dopaminergic neurons and to correlate that with the behavioral effects. Additional cultures and acutely prepared explants were also fixed and stored for histological investigation in order to estimate the loss of dopaminergic neurons in culture and after transplantation. Similar behavioral improvements in terms of significant reductions in amphetamine-induced rotations were observed in rats grafted with FFRT cultures (127%) and rats grafted with cell suspensions (122%), while control animals showed no normalization of rotational behavior. At 84 days after transplantation, there were similar numbers of TH-immunoreactive (TH-ir) neurons in grafts of cultured tissue (775 +/- 98, mean +/- SEM) and grafts of fresh, dissociated cell suspension (806 +/- 105, mean +/- SEM). Cell counts in fresh explants, 7-day-old cultures, and grafted cultures revealed a 68.2% loss of TH-ir cells 7 days after explantation, with an additional 23.1% loss after grafting, leaving 8.7% of the original number of TH-ir cells in the intracerebral grafts. This is to be compared with a survival rate of 9.1% for the TH-ir cells in the cell-suspension grafts. Immunostaining for the calcium-binding proteins calretinin, calbindin, and parvalbumin showed no differences in the neuronal expression of these proteins between the two graft types. In conclusion, we found comparable dopaminergic cell survival and functional effects of tissue-culture grafts and cell-suspension grafts, which currently is the type of graft most commonly used for experimental and clinical grafting. In this sense the result is promising for the development of an effective in vitro storage of fetal nigral tissue, which at the same time would allow neuroprotective and neurotrophic treatment prior to intracerebral transplantation.

Amphetamine↗

Sister chromatid exchanges induced in vitro and in vivo by an extract of black pepper.

Black pepper is a spice widely used in human food. The aim of this investigation was to determine whether an alcoholic extract of the mature berries of black pepper induced genotoxic damage in vivo and in vitro. The first aspect was evaluated in mouse bone marrow cells and the second one in human lymphocytes. In both cases the rate of sister chromatid exchange (SCE) and the replicative index were determined. For the in vivo assay, ip doses of 7.0, 14.0, 28.0 and 56.0 mg/kg body weight were tested, with the following results: (1) a significant increase of SCE frequency in all doses tested compared with the control level (the highest dose produced almost a duplication of the basal rate of SCEs); (2) a similar pattern with regard to cell proliferation kinetics at all doses tested, without significant differences between them. For the in vitro assay, doses of 25.0, 50.0, 75.0 and 100.0 micrograms/ml were tested, with the following results: (1) a significant increase in the frequency of SCEs at all doses tested; a linear regression analysis of the data produced a correlation coefficient of 0.98; (2) a significant reduction in the replicative index, at the two high doses. These results demonstrated that the extract of black pepper was genotoxic in both systems.

Adult↗

A new method of synthesizing biopolymeric affinity ligands.

(1) A new concept for producing soluble polymeric affinity ligands is proposed and exemplified. By solid-phase synthesis, an insoluble hydrophilic polymer is converted into an affinity gel. The gel is hydrolytically degraded to water-soluble affinity polymeric ligands which are recovered and purified. (2) A water-soluble biopolymeric metal-affinity carrier based on an iminodiacetic acid (IDA) derivative of dextran has been synthesized through the modification of Sephadex G-200 by IDA, followed by hydrolysis with dextranase and size-exclusion-chromatographic purification of the high-molecular-mass fragments. (3) The molecular size of the soluble products as a function of hydrolysis time with dextranase from Penicillium sp. was determined. The range of molecular size of the biopolymeric chelating ligand varies from around 200 Da to greater than 580 kDa. (4) The influence of three metal ions chelated with the Sephadex derivative on the hydrolysis rate and the molecular-size distribution of end products was studied. Eu3+ was found to improve the rate of solubilization. Ni2+ and Cu2+ decreased the hydrolysis rate, as compared with that of the metal-free IDA-Sephadex. (5) The method introduced here has the potential of being developed and applied as a general technology for synthesis of soluble multifunctional affinity ligands. Such ligands should be useful for liquid-phase extraction as well as for the synthesis of adsorbents with localized multiple binding sites. Other possible fields of applications are to be found in medicine, where they could be used for slow drug delivery or detoxification, and in analytical chemistry, where they could be used in various assays.

Affinity Labels↗

Sensitivity and specificity of in situ bacterial chain reaction (BCR) in detecting sparse human tumor cells in peripheral blood.

We report here the development of bacterial chain reaction (BCR), a system using micro organisms as nanodevices to amplify and visualize signals from molecular bioprobes such as antibodies, binding proteins, lectins, and oligonucleotides. Unlike conventional enzyme-linked amplification systems in which the amount of enzyme is a constant parameter, in the BCR an enzyme (penicillinase) is used to trigger a proliferative chain reaction producing an exponential increase in enzyme. The detection limits and specificity of BCR were determined using a model system designed to detect and enumerate MCF-7 (a human breast adenocarcinoma cell line) cells disseminated at extremely low frequency (e.g., one tumor cell per million normal cells) among monocluclear cells (MNCs) of human peripheral blood. Results of testing 83 specimens of peripheral blood from presumably healthy donors showed 97.6% specificity. The system was capable of detecting tumor cells at a frequency of 2 x 10(-7).

Adenocarcinoma↗

Treatment of non-Hodgkin's lymphoma of Waldeyer's ring: radiotherapy versus chemotherapy versus combined therapy.

Treatment of stage IA non-Hodgkin's lymphoma (NHL) of Waldeyer's ring remains controversial, probably because of the small number of patients and the scarcity of controlled studies. Between 1981 and 1991, 316 patients with stage I NHL of Waldeyer's ring were randomised for treatment with radiotherapy alone (extended fields), 101 patients; combined chemotherapy with a regimen of CHOP (cyclophosphamide, vincristine, doxorubicin, and prednisone) or CHOP-like (epirubicin instead of doxorubicin), 106 patients; and combined therapy (radiotherapy followed by the same combination chemotherapy), 109 patients. Median follow-up was 6.8 years. Complete response was achieved in 93, 87 and 97%, respectively. Relapses were least frequent in patients treated with combination therapy. The 5-year rate for failure-free survival was 48% for radiation therapy, 45% for the patients who were treated with chemotherapy, which was statistically significantly less than the 83% for patients treated with combined therapy (P < 0.001). Overall survival was also better in the combined therapy arm: 90%, statistically different to 58% for the patients treated with chemotherapy alone and 56% for patients treated with radiation therapy (P < 0.001). Toxicity was mild and late side-effects were not observed in any patients. From these results combined therapy should be considered as the best therapeutic approach in patients with localised NHL of Waldeyer's ring.

Adult↗

Interferon alpha 2b as maintenance therapy in low grade malignant lymphoma improves duration of remission and survival.

We assessed the efficacy and toxicity of interferon alpha 2b (IFN) as maintenance therapy in patients with low grade malignant lymphoma. Between March 1986 and December 1989, 98 patients with low-grade malignant lymphoma in complete remission after conventional chemotherapy were randomly assigned to received IFN, 5.0 MU three times a week for one year, as maintenance therapy (n = 48), or to receive no treatment (control group, n = 50). In March 1994, the median duration of response had not yet been reached in the patients treated with IFN compared to 46 months in the control group. At 9-years 62% of the patients in the IFN arm remain in first complete remission compared to only 25% in the control group (p <.001). In addition, the median duration of survival has not yet been reached in either the IFN arm compared to 74 months in the control group (p <.001). Quality of life was excellent in both groups and severe side effects secondary to IFN treatment were not observed. All patients completed the planned dose of IFN. We conclude that IFN as maintenance therapy in low-grade malignant lymphoma is an excellent therapeutic option because it improves the duration of remission and survival without producing severe side effects or reducing the quality of life.

Adult↗

Technetium-99m-pertechnetate uptake by intrathyroidal parathyroid adenoma.

A 57-yr-old woman was referred for preoperative scintigraphic localization of a parathyroid adenoma. Double-phase 99mTc-sestamibi imaging was performed followed by a separate-day [99mTc]pertechnetate thyroid scan. There was marked accumulation of both tracers by a right lower pole ¿thyroid' nodule which, at surgery, proved to be an intrathyroidal parathyroid adenoma. Hypervascularity is the presumed explanation for rare cases of pertechnetate-avid parathyroid adenomas.

Adenoma↗

Preoperative localization of a parathyroid adenoma with Tc-99m sestamibi imaging in a patient with concomitant nontoxic multinodular goiter.

Preoperative localization of parathyroid adenomas is useful in patients undergoing parathyroidectomy. The authors report a patient with hyperparathyroidism and an associated nontoxic nodular goiter. Technetium-99m sestamibi scintigraphy localized and differentiated the parathyroid adenoma from the multinodular goiter. Excellent correlation was found between preoperative radionuclide imaging and surgical pathology. Technetium-99m sestamibi parathyroid imaging may be a simple and helpful test in the surgical management of parathyroid adenomas, particularly in patients with nontoxic multinodular goiter.

Adenoma↗

Mammary phenotypic expression induced in epidermal cells by embryonic mammary mesenchyme.

The goal of this research was to establish methods for inducing mammary epithelial differentiation from nonmammary epithelium. For this purpose, mid-ventral or dorsal epidermis (skin epithelium; SKE) from 13-day rat or mouse embryos was associated with 13-day embryonic mouse mammary mesenchyme (mammary gland mesenchyme; MGM) (mouse MGM+rat or mouse SKE). The resultant MGM+SKE recombinants as well as controls (homotypic mouse mammary recombinants, homotypic mouse skin recombinants and mouse mammary mesenchyme by itself) were grafted under the renal capsule of syngeneic or athymic female nude mouse hosts. Most female hosts were induced to undergo lactogenesis by grafting an adult pituitary which elicited a state of hyperprolactinemia. Tissue recombinants of mouse MGM+rat or mouse SKE grown for 1 month in vivo formed a hair-bearing keratinized skin from which mammary ductal structures extended into the mesenchyme. The ducts were composed of columnar luminal epithelial cells as well as basal, actin-positive myoepithelial cells. When grown in pituitary-grafted hosts, the ductal epithelial cells expressed casein and alpha-lactalbumin as judged by immunocytochemistry. The expression of caseins in MGM+SKE recombinants was confirmed by Western blot. The epithelial cells in mouse MGM+rat SKE recombinants expressing milk proteins were shown to be rat cells while the surrounding connective tissue was composed of mouse cells based upon staining with Hoechst dye 33258. Using mammary-specific markers, these studies confirmed the earlier morphological studies of Propper and unequivocally demonstrated for the first time that embryonic mammary mesenchyme can induce morphological and functional mammary differentiation from nonmammary epithelium.

Animals↗

Transgenes expressing the Wnt-1 and int-2 proto-oncogenes cooperate during mammary carcinogenesis in doubly transgenic mice.

The Wnt-1 and int-2 proto-oncogenes are transcriptionally activated by mouse mammary tumor virus insertion mutations in virus-induced tumors and encode secretory glycoproteins. To determine whether these two genes can cooperate during carcinogenesis, we have crossed two previously characterized lines of transgenic mice to obtain bitransgenic animals carrying both Wnt-1 and int-2 transgenes under the control of the mouse mammary tumor virus long terminal repeat. Mammary carcinomas appear earlier and with higher frequency in the bitransgenic animals, especially the males, than in either parental line. Nearly all bitransgenic males develop mammary neoplasms within 8 months of birth, whereas only 15% of Wnt-1 transgenic males and none of the int-2 transgenic males have tumors. In virgin bitransgenic females, tumors occur approximately 2 months earlier than in their Wnt-1 transgenic siblings; int-2 transgenic females rarely exhibit tumors. Preneoplastic glands from the bitransgenic animals of either sex demonstrate pronounced epithelial hyperplasia similar to that seen in Wnt-1 transgenic virgin females and males, and both transgenes are expressed in the hyperplastic glands and mammary tumors. RNA from the int-2 transgene is more abundant in mammary glands from bitransgenic animals than from int-2 transgenic animals; the increase is associated with high levels of RNA specific for keratin genes 14 and 18, suggesting that Wnt-1-induced epithelial hyperplasia is responsible for the observed increase in expression of the int-2 transgene.

Animals↗

Developmental response of adult mammary epithelial cells to various fetal and neonatal mesenchymes.

Adult mouse mammary epithelial cells were isolated and grown in combination with mesenchyme from the following sources: embryonic preputial gland, foot skin, tail skin, genital tubercle skin, mammary gland, and neonatal uterus, vagina and urinary bladder. Following 1 month of in vivo cultivation of the tissue recombinants as grafts underneath the renal capsule of normal female or hyperprolactinaemic (pituitary-grafted) hosts, the specimens were analysed histologically and immunocytochemically for the expression of milk proteins, smooth muscle actin, and cytokeratins. Mesenchymal effects on adult mammary epithelium varied with the source of the mesenchyme and the hormonal status of the host. In normal female hosts preputial gland mesenchyme induced extensive mammary epithelial growth and ductal branching morphogenesis with epithelial differentiation and ductal pattern being comparable to that observed in homotypic recombinants composed of mammary gland mesenchyme plus adult mammary epithelium. Other mesenchymes (from foot, tail, genital tubercle and uterus) preserved ductal morphology and normal epithelial differentiation, but elicited minimal epithelial growth and branching morphogenesis in adult mammary epithelium. In association with urogenital sinus, vaginal or bladder mesenchymes ductal branching morphogenesis of the mammary epithelium was absent or greatly distorted and the epithelium exhibited a stratified cuboidal phenotype even though considerable epithelial growth had occurred. In hyperprolactinaemic hosts (which received a pituitary graft) epithelial growth, alveolar morphogenesis, and synthesis of casein and milk fat globule protein was stimulated in all tissue recombinants although to different extents. Alveolar morphogenesis and milk protein expression were extensive in tissue recombinants prepared with mesenchyme from embryonic mammary gland, preputial gland, tail skin and urinary bladder, but were minimal in tissue recombinants prepared with foot or genital tubercle skin. Few milk-protein-positive alveoli formed in tissue recombinants composed of mammary epithelium combined with urogenital sinus or vaginal mesenchyme, even following growth in pituitary-grafted hosts. These findings demonstrate: (1) that adult mammary epithelial cells are responsive to the growth-promoting influences of heterotypic embryonic and neonatal mesenchymes; (2) that mammary growth and branching morphogenesis are induced to variable extents by different mesenchymes; (3) that fibrous (non-adipose) mesenchymes are effective inducers of mammary epithelial development; and (4) that the ability to form alveoli and produce milk proteins in adult mammary epithelial cells is critically dependent upon the nature of the connective tissue environment.

Animals↗

Etiology of prosthetic anastomotic false aneurysms: pathologic and structural evaluation in 26 cases.

To determine the etiology of anastomotic false aneurysms (AFAs), 26 textile graft specimens, removed because of AFA, were studied morphologically, histologically and by scanning electron microscopy. No cases of suture-related failures leading to AFA were found. Nine cases of frayed grafts were documented but were not the cause of AFA formation. In three cases, chemical degradation of the fibres, which may have been secondary to lipid infiltration, may have contributed to AFA formation. There were no cases of overt clinical infection, but the presence of bacteria was documented by scanning electron microscopy in 20 cases. The role of bacteria is not well defined, but they may be a factor in host arterial-wall degeneration as a cause for AFA formation.

Adult↗

[Hodgkin's disease treated with chemotherapy at reduced dosage].

Fifty-six patients with Hodgkin's disease in advanced stages (III-IV) were entered in a study aimed to compare standard doses (28 cases) with low doses (28 cases) ABVD regimen. Both groups had similar clinico-pathological features. The group receiving standard doses chemotherapy achieved complete remission rate of 89% versus 85% for the low-doses group. The number of relapses was, respectively, 8 and 9, and the duration of remission was 66 and 57 months, respectively. Presently, 24 patients have died, 11 of the standard doses group and 13 of the reduced doses group. Toxicity was notably higher in the group receiving standard dosage, where fatal myelosuppression was present in two instances. Those data appear to suggest that chemotherapy with reduced doses for Hodgkin's disease can attain similar response rate and survival than those achieved with standard treatment, with the advantages of a lower toxicity.

Antineoplastic Combined Chemotherapy Protocols↗

Odontodysplasia in a pediatric patient: literature review and case report.

A case of a three-year-old Caucasian male with odontodysplasia is presented. Although the etiology of this condition is unknown, this anomaly involves both the mesodermal and ectodermal dental components and results in deficient and abnormal formation of dentin and enamel. The orofacial characteristics and dental findings of the condition are presented in this case report.

Child, Preschool↗

Human umbilical vein grafts as infrainguinal bypasses: long-term clinical follow-up and pathological investigation of explanted grafts.

This retrospective study examines 105 consecutive infrainguinal bypasses using human umbilical vein (HUV) grafts as blood conduits over a 9 year period. In addition, 39 segments of these grafts were harvested at reoperation and submitted for morphologic, histologic, and scanning electron microscopy examinations. Fifty-two below-knee femoro-popliteal and 53 femoro-distal bypasses were performed in 93 patients. Sixty-seven per cent of these bypasses were performed for either rest pain (18%) or gangrene (49%) and 29% were performed for acute ischemia. Primary and secondary patency rates were not statistically different and were 38% and 29% at 1 and 3 years respectively. Factors found to have a significant effect on patency were site of distal anastomosis, state of distal run-off, and indication for operation. Other complications requiring reoperation included aneurysm formation (8 cases), infection (6 cases), mural thrombus (2 cases), and stenosis (2 cases). Pathologic examination revealed a damaged luminal surface in 22 grafts, deep folds in 8 grafts and delamination of the graft wall in 16 cases. Bacteria were seen in the folds of the grafts and, in addition, the presence of bacteria was documented in 7 out of 26 clinically non-infected grafts. The combination of poor patency rates and pathologic evidence of biodegradative phenomena have led us to discontinue HUV as an arterial substitute.

Adult↗

Femorodistal bypass using the chemically processed human umbilical vein graft: 9-year experience.

To evaluate patency, limb salvage rates and complications associated with the use of human umbilical vein as an arterial substitute, the authors carried out a retrospective review of 52 femorodistal bypasses performed over 9 years using human umbilical vein. Indications for operation included acute ischemia, rest pain and nonhealing ulceration or gangrene. There were 28 early occlusions, which resulted in a 1-month primary patency rate of 46% and a secondary patency rate of 52%. The primary and secondary patency rates at 1 year were 18% and 19% respectively. The mean limb salvage rate at 1 month was 57% and at 1 year 34%. Three aneurysms occurred (two anastomotic, one graft). Factors found to have a significant effect on patency rates were indication for operation and state of distal runoff. Infection occurred in 6% of grafts and led to amputation in every case. These disappointing results have caused the authors to discontinue use of human umbilical vein as an arterial substitute.

Adult↗