O objetivo do presente estudo foi avaliar microscopicamente a resposta inflamatória do tecido conjuntivo subcutâneo de ratos quando em contato com formulações à base de própolis (A70D e D70D). Foram utilizados 15 ratos machos da linhagem Wistar, com aproximadamente 40 dias de vida. Em períodos experimentais de 7, 15 e 30 dias, foram implantados tubos de polietileno contendo materiais diferentes, formando 4 grupos: Grupo I - Ca(OH)2, Grupo II - A70D, Grupo III - D70D, Grupo IV Controle (tubo vazio). Após os períodos experimentais, os animais foram mortos e os tecidos contendo os implantes foram removidos para processamento histotécnico. Os cortes microscópicos semi-seriados de 5µm de espessura foram corados pela técnica da Hematoxilina e Eosina (H.E.). A reação tecidual frente ao material utilizado foi analisada de forma semi-quantitativa e qualitativa. Para análise estatística foi utilizado o teste de Kruskal-Wallis com p<0,05. Os resultados mostraram intensa reação inflamatória para todas as pastas aos 7 dias, com intensa proliferação angioblástica, moderado infiltrado neutrofílico e necrose por coagulação no grupo do hidróxido de cálcio. Com o decorrer dos períodos a intensidade da reação diminuiu, com redução significativa em 30 dias. As pastas A70D e D70D apresentaram resultados semelhantes, porém aos 30 dias a reação se manteve mais intensa para a pasta D70D. Estes resultados sugerem a possibilidade da própolis ser utilizada como medicação intracanal. / The aim of this study was to evaluate the inflammatory tissue response of different based propolis pastes for endodontic use (A70D and D70D). 15 male rats of the Wistar strain, approximately 40 days of life were used. Polyethylene tubes containing different materials were divided in 4 groups: Group I - Ca(OH)2, Group II - A70D, Group III - D70D, Group IV - Control (empty tube ). The inflammatory response of subcutaneous tissue was analyzed after 7, 15 and, 30 days. After the experimental periods, animals were killed and tissues containing the implants were removed and prepared using conventional histological procedures. Microscopic semi-serial sections of 5m in thickness were stained by the hematoxylin and eosin technique (HE). The tissue reaction was evaluated using semi-quantitative and qualitative criteria. Statistical analysis was performed using the Kruskal-Wallis test with p <0.05. The results showed intense inflammatory reaction in all the groups at the 7 day period. Intense proliferation of vessels, moderate neutrophilic infiltrate and necrosis by coagulation in the group of calcium hydroxide. The intensity of the reaction decreased with significant reduction after 30 days. The A70D and D70D pastes showed similar results, but at the 30 periods the most intense reaction was found in the D70D paste. These results suggest the possibility that propolis can be used as an intracanal medication.

Activated Heat Shock Transcription Factor 1 (HSF1) has received attention in recent literature as a therapeutic effector in diseases of protein misfolding, as an immune modulating adjuvant in tumor regression, and as a trigger for gene therapy transcription. In its normal function, activated HSF1 enhances heat shock protein (Hsp) expression when additional molecular chaperoning is required (i.e., in situations of proteotoxic stress, including thermal stress) in a process known as the heat shock (HS) response. Thus, HSF1 acts as an environmental sensor, and a harness based on the biology of this capability enables transcription of genes for engineered purposes. The hypothesis of this thesis is that a harness of the heat shock response, when paired with a therapeutic mechanism, will refine novel therapies. Extensions to the concept of deliberately activating HSF1's normal functions for therapeutic purposes are examined through in vitro trials and in vivo preliminary studies that feature the use of HSF1 as a regulator of therapy. Successful in vitro work translated to pioneering preclinical studies, launched at the University of Florida's Center for Environmental and Human Toxicology. Collaboration supported the development of an innovative project to treat solid tumors using a recombinant virus system. The system was designed to facilitate intratumoral delivery of a previously characterized molecular switch, which was newly engineered to control cytotoxic gene transcription that produced dramatic consequences in cells of human origin. Central to the targeting of the in vivo therapy, is a transient, initial trigger: a thermal dose, delivered to solid tumors, which localizes HSF1 activation (a constitutively active mouse HSF1 construct was also produced to aid clarification of physiological consequences associated with deliberately upregulating HSF1 activity in vivo). Gene transcription was expected to ensue to both cause and sustain tumor regression through other regulatory elements of the molecular switch. Results demonstrated practical potential to achieve a therapeutic outcome of solid tumor regression and define contemporary challenges that continuing research directions (e.g.: production of additional viral vectors, an improved animal model, and a refined heat system) now confront in order to target and safely regulate even more potent, novel therapeutic agents.

We hypothesized that slip point of subcutaneous (s.c.) adipose tissue lipids would predict beef carcass quality. To address our hypothesis, 79 M. longissimus dorsi (LD) steaks from cattle of unknown background were used to provide information on slip points, percentage intramuscular lipid, fatty acid composition, and MUFA:SFA ratios. Overlying s.c. adipose tissue was separated from the muscle lean, which contained intramuscular (i.m.) adipose tissue. Lipids were extracted from s.c. adipose tissue and muscle lean by a modified chloroform:methanol procedure and subjected to various analyses. The hypothesis was tested by developing regression equations to determine which fatty acid variables were most useful in predicting carcass composition. There was a high correlation between s.c. MUFA:SFA ratio and s.c. slip points (P < 0.001) with an R2 of 0.557. Also, the MUFA:SFA fatty acid ratios of s.c. and i.m. adipose tissue were significantly correlated and an R2 of 0.440 was observed (P < 0.001) when regressed against each other. The current data set observed s.c. MUFA:SFA ratios (0.73) lower than previous studies, which suggests a population of young or unfinished cattle. This study demonstrated that it is possible to predict the intramuscular lipid (IML) MUFA:SFA ratio by measuring s.c. slip point (R2 of 0.097; P < 0.01). However, our hypothesis of predicting amount of marbling, hence quality grade, from the melting temperature of s.c. adipose tissue lipids proved incorrect (R2 = 0.001). Nonetheless, these data indicate that LD fatty acid composition can be estimated by measuring s.c. adipose tissue slip point.

Delivery of protein therapeutics is restricted to intravenous infusions due to protein-dependent problems including low solubilities, high viscosities, and physical instabilities. The ability to inject high concentrations of proteins via subcutaneous injections would increase accessibility and compliance. Large particles of a protein in a non-aqueous solvent can decrease the viscosity over a solution of equally concentrated individual protein molecules. The lower viscosity of a particle suspension is due to decreased surface area resulting in reduced electroviscous effects, solvation and deviations of the particle shape from a spherical geometry. Additional studies show that aqueous-based dispersions of antibody nanoclusters can be formed by increasing the attractive interactions between protein molecules using the excluded volume effects of extrinsic crowding agents. These novel, equilibrium, nanoclusters are maintained by a balance of highly attractive interactions and weak electrostatic repulsive interactions near the protein’s pI. These protein nanoclusters are ideal for subcutaneous delivery as they have low interactions between the colloids, are reversible in nature, and dissolve rapidly upon dilution in a buffer media. Through in vivo mouse studies, the bioavailability of a monoclonal antibody in the dispersion is prolonged and higher doses can be administered versus a solution. Overall, these studies with high concentration, low viscosity subcutaneous injections of protein therapeutics open new opportunities in biotechnology. For oral delivery of itraconzole, controlled flocculation of individual polymerically-stabilized nanoparticles is used to increase supersaturation. Flocculation of these nanoparticles is achieved by desolvating the polymer by changing the pH. The flocculated dispersions can then be easily filtered. The final amorphous powder maintains high supersaturation with simulated stomach and small intestine conditions and improves bioavailability of itraconazole, over the commercial product, Sporanox®. / text

The MicroImager is a fast, high resolution, real time, digital autoradiographic imaging tool with broad applications. This study utilizes the MicroImager to evaluate radiolabeled drug behavior in subcutaneous tissue. Experiments were conducted in conjunction with mathematical models to determine the diffusion coefficient (D) and elimination constant (k) for radiolabeled dexamethasone. Osmotic pumps containing [3H]dexamethasone were implanted into rat subcutaneous tissue over 6h, 24 h, and 60 h. Local tissue was explanted and slides were prepared for imaging. The MicroImager was then used to quantify the local concentration of 3H-dexamethasone in the tissue surrounding the tip of the osmotic pump. Betavision+ software was used to obtain local concentration profiles. These were then compared to a mathematical model to determine the diffusion coefficient and elimination constant for the radiolabeled drug. The diffusion coefficient for dexamethasone in rat subcutaneous tissue is 4.11 ± 1.77 x 10-10 m2/s. The elimination constant is 3.65 ± 2.24 x 10-5 s-1.A similar experiment was conducted to determine the diffusion coefficient through different means. [3H]dexamethasone was injected into the rat subcutaneous tissue for a 2.5 min and a 20 min period. A different mathematical model was applied and the diffusion coefficient was found to be 4.01 ± 2.01 x 10-10 m2/s.

Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2006. / Härtill 4 uppsatser.

O objetivo do presente estudo foi avaliar microscopicamente a resposta inflamatória do tecido conjuntivo subcutâneo de ratos quando em contato com formulações à base de própolis (A70D e D70D). Foram utilizados 15 ratos machos da linhagem Wistar, com aproximadamente 40 dias de vida. Em períodos experimentais de 7, 15 e 30 dias, foram implantados tubos de polietileno contendo materiais diferentes, formando 4 grupos: Grupo I - Ca(OH)2, Grupo II - A70D, Grupo III - D70D, Grupo IV Controle (tubo vazio). Após os períodos experimentais, os animais foram mortos e os tecidos contendo os implantes foram removidos para processamento histotécnico. Os cortes microscópicos semi-seriados de 5µm de espessura foram corados pela técnica da Hematoxilina e Eosina (H.E.). A reação tecidual frente ao material utilizado foi analisada de forma semi-quantitativa e qualitativa. Para análise estatística foi utilizado o teste de Kruskal-Wallis com p<0,05. Os resultados mostraram intensa reação inflamatória para todas as pastas aos 7 dias, com intensa proliferação angioblástica, moderado infiltrado neutrofílico e necrose por coagulação no grupo do hidróxido de cálcio. Com o decorrer dos períodos a intensidade da reação diminuiu, com redução significativa em 30 dias. As pastas A70D e D70D apresentaram resultados semelhantes, porém aos 30 dias a reação se manteve mais intensa para a pasta D70D. Estes resultados sugerem a possibilidade da própolis ser utilizada como medicação intracanal. / The aim of this study was to evaluate the inflammatory tissue response of different based propolis pastes for endodontic use (A70D and D70D). 15 male rats of the Wistar strain, approximately 40 days of life were used. Polyethylene tubes containing different materials were divided in 4 groups: Group I - Ca(OH)2, Group II - A70D, Group III - D70D, Group IV - Control (empty tube ). The inflammatory response of subcutaneous tissue was analyzed after 7, 15 and, 30 days. After the experimental periods, animals were killed and tissues containing the implants were removed and prepared using conventional histological procedures. Microscopic semi-serial sections of 5m in thickness were stained by the hematoxylin and eosin technique (HE). The tissue reaction was evaluated using semi-quantitative and qualitative criteria. Statistical analysis was performed using the Kruskal-Wallis test with p <0.05. The results showed intense inflammatory reaction in all the groups at the 7 day period. Intense proliferation of vessels, moderate neutrophilic infiltrate and necrosis by coagulation in the group of calcium hydroxide. The intensity of the reaction decreased with significant reduction after 30 days. The A70D and D70D pastes showed similar results, but at the 30 periods the most intense reaction was found in the D70D paste. These results suggest the possibility that propolis can be used as an intracanal medication.

Subcutaneous (SC) injection is a safe and effective way to administrate monoclonal antibodies (mAbs) and very often valued by patients and healthcare providers alike. A SC injection offers noteworthy advantages over intravenous (IV) injections, such as lowering hospital and clinical costs. Additionally, SC injections requires a lower number administration and allows for self-administrated or caregiver-supported dosing at home, thus a reduced healthcare resource utilization and healthcare provider time. In recent years biotherapeutics have been one of the fastest growing class of drugs in the pharmaceutical industry due to effective therapeutics with target specificity and potency. Biopharmaceuticals are usually administrated through IV, intramuscular (IM) or SC injection. However, patients are not administrated drugs, they are administrated formulations containing a drug. It is also important to recognize that biopharmaceuticals that are meant for SC injection are usually formulated at an acidic (4-6) pH, which can be attained by using a variety of excipients and stabilising agents. Excipients such as sugar, salt and surfactant are commonly found in biopharmaceuticals and can be used to attain a multi-year shelf life. Additionally, biopharmaceuticals can form aggregates and this occurrence happens due to loss of a stabilizing excipient. Hence the importance of understanding how and why different excipients are or can be used in a formulation.

Objective: This study was undertaken to determine the most appropriate management of the subcutaneous tissue of midline vertical incisions with 3 cm or more of subcutaneous fat. Study design: Patients undergoing surgery within the Division of Gynecologic Oncology at University of South Florida and East Tennessee State University with 3 cm or more of subcutaneous fat were randomly assigned to 1 of 3 groups: suture approximation of Camper's fascia, closed suction drainage of the subcutaneous space, or no intervention as a control group. Participants were evaluated daily during postoperative hospitalization and at 2 and 6 weeks postoperatively as an outpatient. Demographic information, perioperative data, and wound complications were recorded and then analyzed with χ2, t test, analysis of variance, and logistic regression where appropriate. Results: Two hundred twenty-five patients were enrolled with 222 eligible for evaluation. Wound complications were observed in 34 (15.3%) patients, and 25 of these women also had wound disruption. Overall wound complication and wound disruption rates were not significantly different between groups: suture (12.8%, 7.7%), drain (17.9%, 14.9%), control (15.6%, 11.7%); P = .70 and P = .39, respectively. Conclusion: Suture approximation or drainage of the subcutaneous tissues of women with 3 cm or more subcutaneous fat measured in midline vertical incisions resulted in no significant change in the incidence of overall wound complications or superficial wound disruption.

We measured black bear (Ursus americanus) reproduction and cub survival during 1994 - 1998, and 1995 - 1999, respectively, in the George Washington and Jefferson National Forests in Virginia to determine age-specific and overall cub production and cub survival. We observed females in estrus between 6 June and 22 August; the mean date of estrus was 17 July. Ages of primiparity ranged between 3 and 5 years with an average of 3.36 years (n=11, SE=0.15). Average litter size for 1995 - 1998 was 2.32 cubs/litter (SE=0.11, n=53) and 85.7% of available females &ge; age 4 (those not accompanied by cubs) reproduced in a given den season. We monitored 98 (48M:50F) black bear cubs equipped with expandable radio-collars (Higgins 1997) or radio transmitters implanted subcutaneously between 1995 and 1999 to estimate cub survival. Kaplan-Meier staggered entry analysis provided 306-day survival rates for 82 cubs. The survival estimates for males and females were 73% (0.49, 0.96) and 91% (0.80, 1.00), respectively. The overall 306-day survival rate for all cubs was 81% (0.67,0.94) using Kaplan-Meier and 76% (0.63, 0.92) using Heisey-Fuller (Mayfield) methods. We also evaluated the utility of radio transmitters implanted subcutaneously in 42 (21M:21F) wild black bear (Ursus americanus) cubs from 2 study areas in Virginia between 1996 and 1999 to monitor first year cub survival. More than 64% (27 of 42) of the implants fell out prematurely (2-198 days), and 16.6% (7 of 42) failed for unknown reasons. Less than 5% (2 of 42) of these cubs denned wearing failed implants, and 9.5% (4 of 42) experienced mortality less than 1 month after implant surgery. About 9.5% (4 of 42) of implanted black bear cubs wore working transmitters through to the following den season. / Master of Science