Iceberg lettuce cultivated in different systems of planting and sources of fertilizer / Alface americana cultivada em diferentes sistemas de plantio e fontes de fertilizantes

Organic fertilization is a cheaper and highly effective option for profitability and consequent improvement of the soil's physical, chemical, and biological structure. Thus, the objective of this work was to evaluate different types of fertilization: organic (poultry shed litter), mineral, and leaf path on yield parameters of lettuce grown in various types of planting. The treatments consisted of using two planting systems (P1 - Line and P2 - quincunxes) and mineral and organic fertilizers (A1 - mineral fertilization; A2 - mineral fertilization + leaf fertilization; A3 - organic fertilization with poultry shed litter and A4 - fertilization organic + mineral). The experimental units consisted of 36 and 52 plants, respectively, for treatments P1 and P2, and all central plants of the experimental unit were evaluated. Heart height, fresh mass, and leaf number were observed. The mineral and mineral + leaf treatments did not differentiate, either in line or in quincunxes. The treatment that stood out about the analyzed variables was the organic fertilization and quincunxes planting system, reflecting a more significant number of lettuce plants and better use of the area.

A adubação orgânica é uma opção mais barata e de grande eficácia em relação à rentabilidade e consequente melhoria da estrutura física, química e biológica do solo. Desta forma, o objetivo deste trabalho foi avaliar diferentes tipos de adubação: orgânica (cama de frango), mineral e via foliar sobre parâmetros de produtividade de alface cultivada em diferentes tipos de plantio. Os tratamentos consistiram na utilização de dois sistemas de plantio (P1 - Linha e P2 - Quincôncio) e adubações minerais e orgânicas (A1 - adubação mineral; A2 - adubação mineral + adubação foliar; A3 - adubação orgânica com cama de aviário e A4 - adubação orgânica + mineral). As unidades experimentais foram compostas por 36 e 52 plantas, respectivamente, para os tratamentos em linha e em quincôncio, sendo avaliadas todas as plantas centrais da unidade experimental. Foram observados a altura do coração, massa fresca e número de folhas. Os tratamentos mineral e mineral + foliar não diferenciaram entre si, tanto em linha quanto em quincôncio. O tratamento que se destacou em relação às variáveis analisadas foi aquele baseado na adubação orgânica e sistema de plantio em quincôncio, refletindo em maior número de pés de alface e melhor aproveitamento da área.

Iceberg lettuce cultivated in different systems of planting and sources of fertilizer

Abstract Organic fertilization is a cheaper and highly effective option for profitability and consequent improvement of the soil's physical, chemical, and biological structure. Thus, the objective of this work was to evaluate different types of fertilization: organic (poultry shed litter), mineral, and leaf path on yield parameters of lettuce grown in various types of planting. The treatments consisted of using two planting systems (P1 - Line and P2 - quincunxes) and mineral and organic fertilizers (A1 - mineral fertilization; A2 - mineral fertilization + leaf fertilization; A3 - organic fertilization with poultry shed litter and A4 - fertilization organic + mineral). The experimental units consisted of 36 and 52 plants, respectively, for treatments P1 and P2, and all central plants of the experimental unit were evaluated. Heart height, fresh mass, and leaf number were observed. The mineral and mineral + leaf treatments did not differentiate, either in line or in quincunxes. The treatment that stood out about the analyzed variables was the organic fertilization and quincunxes planting system, reflecting a more significant number of lettuce plants and better use of the area.

Resumo A adubação orgânica é uma opção mais barata e de grande eficácia em relação à rentabilidade e consequente melhoria da estrutura física, química e biológica do solo. Desta forma, o objetivo deste trabalho foi avaliar diferentes tipos de adubação: orgânica (cama de frango), mineral e via foliar sobre parâmetros de produtividade de alface cultivada em diferentes tipos de plantio. Os tratamentos consistiram na utilização de dois sistemas de plantio (P1 - Linha e P2 - Quincôncio) e adubações minerais e orgânicas (A1 - adubação mineral; A2 - adubação mineral + adubação foliar; A3 - adubação orgânica com cama de aviário e A4 - adubação orgânica + mineral). As unidades experimentais foram compostas por 36 e 52 plantas, respectivamente, para os tratamentos em linha e em quincôncio, sendo avaliadas todas as plantas centrais da unidade experimental. Foram observados a altura do coração, massa fresca e número de folhas. Os tratamentos mineral e mineral + foliar não diferenciaram entre si, tanto em linha quanto em quincôncio. O tratamento que se destacou em relação às variáveis analisadas foi aquele baseado na adubação orgânica e sistema de plantio em quincôncio, refletindo em maior número de pés de alface e melhor aproveitamento da área.

Iceberg lettuce cultivated in different systems of planting and sources of fertilizer.

Organic fertilization is a cheaper and highly effective option for profitability and consequent improvement of the soil's physical, chemical, and biological structure. Thus, the objective of this work was to evaluate different types of fertilization: organic (poultry shed litter), mineral, and leaf path on yield parameters of lettuce grown in various types of planting. The treatments consisted of using two planting systems (P1 - Line and P2 - quincunxes) and mineral and organic fertilizers (A1 - mineral fertilization; A2 - mineral fertilization + leaf fertilization; A3 - organic fertilization with poultry shed litter and A4 - fertilization organic + mineral). The experimental units consisted of 36 and 52 plants, respectively, for treatments P1 and P2, and all central plants of the experimental unit were evaluated. Heart height, fresh mass, and leaf number were observed. The mineral and mineral + leaf treatments did not differentiate, either in line or in quincunxes. The treatment that stood out about the analyzed variables was the organic fertilization and quincunxes planting system, reflecting a more significant number of lettuce plants and better use of the area.

Incidence and risk factors of injuries in Brazilian elite handball players: A prospective cohort study.

The aim of this study was to investigate the incidence and risk factors for handball injuries in Brazilian elite handball players. Overall, 339 athletes from 21 handball teams who participated in the two main Brazilian championships were followed up during a season. In total, 312 injuries were reported by 201 athletes. The injury incidence rate during training was 3.7/1000 h, and during matches was 20.3/1000 matches. Ankle (19.4%, n = 46) and knee (13.5%, n = 32) were the body regions most affected by traumatic injuries. Shoulders (44.0%, n = 33) and knee (26.7%, n = 20) were the body regions most affected by overuse injuries. Muscle injuries (27.1%, n = 68) was the traumatic injury type most reported. Tendinopathy (91.8%, n = 56) was the overuse injury type most observed. Previous injury (OR: 2.42, CI 95%: 1.51-3.89) and an additional match per week (OR: 1.31, CI 95%: 1.05-1.62) were associated with a higher risk of overuse injury. Female athletes (OR: 1.56, CI 95%: 1.08-2.25) and an additional hour of training per week (OR: 1.09, CI 95%: 1.02-1.15) were associated with a higher risk of traumatic injury. This study showed that athletes with previous injury have shown a high risk of developing an overuse injury.

Genetic divergence among sweet corn lines estimated by microsatellite markers.

The purpose of this study was to analyze the genetic diversity of 15 sugary-1 sweet corn lines by microsatellite markers. One hundred pairs of simple sequence repeat primers that were mapped for field corn were tested. Of these primers, 15% were polymorphic, and all were selected for the evaluation. These primers identified a total of 39 alleles among the 15 loci that were evaluated. The number of alleles per locus in the genotypes ranged from 2 to 4, with an average of 2.60 alleles per locus; the highest number of alleles was observed at the loci Bnlg1083, Umc1241, and Umc1590. The occurrence of null alleles at locus Umc1363 was evident only in line DN44. The proportion of polymorphic loci was the highest in lines DN17.1 and DN6 (73.33%), whereas lines DN47, DN23, and DN28 were more monomorphic than other lines. The loci Bnlg1083 and Umc1506 were polymorphic in 8 and 7 lines, respectively, indicating that these loci might be effective and promising for the identification of polymorphism in other sweet corn lines. The genetic diversity calculated by Rogers' genetic distances indicated the lowest genetic similarity between lines DN9 and DN28 (0.7603) and the highest similarity between lines DN19 and DN6 (0.3724). The dendrogram obtained by the unweighted pair-group method based on arithmetic averages indicated the formation of 4 major groups, showing the crossing of the genotypes DN19 and DN6 with DN8 as a possible alternative for the expression of heterozygosis.

Intra-operative frozen section analysis for suspected early-stage ovarian cancer: 11 years of Gateshead Cancer Centre experience.

OBJECTIVE: In centres in which intra-operative frozen section (FS) analysis is not performed, 'apparent' early-stage ovarian cancer diagnosed after surgery on paraffin section may require further restaging laparotomy or adjuvant chemotherapy. Previous studies on FS analysis have reported high sensitivity, specificity and overall accuracy. The objective of this article is to present the largest published dataset on the accuracy of FS analysis over an 11-year period from a single institution. DESIGN: Diagnostic test accuracy. SETTING: Northern Gynaecological Oncology Centre and Department of Cellular Pathology, Gateshead, UK. POPULATION: 1439 intra-operative FS analyses performed between January 2000 and December 2010 for suspected ovarian cancer. METHODS: Prospectively collected data on FS analysis were compared with gold standard paraffin section. MAIN OUTCOME MEASURES: Sensitivity, specificity, likelihood ratios and post-test probability. RESULTS: The overall sensitivity and specificity of FS analysis were 91.2% and 98.6%, respectively. Positive and negative likelihood ratios were 64.7% and 0.09%, respectively. The pre-test probability of an ovarian tumour being borderline or malignant was 45.8%. When FS analysis was reported to be positive, the post-test probability increased to 98% (confidence interval, 97-99%). Conversely, when FS analysis was reported to be negative, the post-test probability decreased to 7% (confidence interval, 6-9%). The majority of false test results were either borderline tumours or of mucinous differentiation. CONCLUSIONS: Intra-operative FS analysis has excellent diagnostic test accuracy and assists gynaecological oncologists to perform the appropriate surgery in 95% of cases, thereby preventing the morbidity of surgical staging in benign cases and the morbidity of restaging procedures or chemotherapy in early-stage malignant tumours.

Interventions for reducing anxiety in women undergoing colposcopy.

BACKGROUND: Before cervical cancer develops the cells of the cervix become abnormal. Following an abnormal cervical smear colposcopy is performed. Colposcopy is the visualisation of the cervix using a binocular microscope. Women experience high levels of anxiety and negative emotional responses at all stages of cervical screening. High levels of anxiety before and during colposcopy can have adverse consequences, including pain and discomfort during the procedure and high loss to follow-up rates. This review evaluates interventions designed to reduce anxiety levels during colposcopic examination. OBJECTIVES: To compare the efficacy of various interventions aimed at reducing anxiety during colposcopic examination in women. SEARCH STRATEGY: The Cochrane Central Register of Controlled Trials (CENTRAL), (Cochrane Library, Issue 1, 2006) MEDLINE (1951-2006), EMBASE (1980-2006), CINAHL (1982-2006), Psych Lit and CancerLit, NHMRC Clinical Trials Register, UKCCCR Register of Cancer Trials, Meta-Register and Physician Data Query Protocols. SELECTION CRITERIA: Randomised and quasi randomised controlled trials of interventions to reduce anxiety during colposcopic examination. DATA COLLECTION AND ANALYSIS: One author searched the citations and reference lists. Studies that appeared to meet inclusion criteria were retrieved and assessed independently by the remaining three authors. The methodological quality of included studies was assessed using the Cochrane Collaboration Back Review Group's methodological quality criteria (van Tulder 2003). MAIN RESULTS: Eleven trials were included, these trials used various interventions to reduce anxiety. These examined 1441 women's anxiety levels after different types of intervention. These included: Information leaflets - (proved not to be associated with anxiety reduction). Counselling: pre-colposcopic counselling was not associated with anxiety reduction. Information leaflets and information video and pre-colposcopy counselling was not associated with a reduction in anxiety levels. Listening to music during colposcopy: this intervention was associated with reduction in anxiety levels (p < 0.002). Video colposcopy was associated with reduction in anxiety levels, and the reduction in anxiety was significant (p < 0.0002). Information using graphs and verbal information and information video versus information only when sought: There was no significant reduction in the level of anxiety in the intervention group. Information leaflets and information video versus information leaflets only: There was a reduction in anxiety levels in the intervention group compared to the control group (p < 0.00001). AUTHORS' CONCLUSIONS: Anxiety appears to be reduced by playing music during colposcopy, showing information videos prior to colposcopy and viewing video colposcopy during the procedure. Although information leaflets did not reduce anxiety levels, they did increase knowledge levels and so are useful in obtaining clinical consent to the colposcopic procedure.

Tradução e adaptação cultural do WORC: um questionário de qualidade de vida para alterações do manguito rotador / Translation and cultural adaptation of WORC: a quality-of-life questionnaire for rotator cuff disorders

OBJETIVO: Realizar a tradução e a adaptação cultural do questionário WORC (The Western Ontario Rotator Cuff Index) para a língua portuguesa para ulterior validação no Brasil. MÉTODOS: O protocolo aplicado consistiu em: 1) Preparação, 2) Tradução, 3) Tradução de volta à língua original (retro-tradução), 4) Interrogatório Cognitivo e 5) Relato de Informações. Ao serem concluídas as etapas de tradução e retro-tradução, as versões foram enviadas para os autores do WORC original, que as aprovaram para continuação do estudo. A versão em Português foi aplicada a 35 pacientes com disfunções do manguito rotador para verificar o nível de compreensão do instrumento. A idade média foi 57 anos (DP=13), 63 por cento eram do sexo feminino e 74 por cento tinham nível de escolaridade de primeiro grau incompleto. A versão brasileira final do WORC foi definida após se conseguir menos que 15 por cento de "não compreensão" em cada item. Para análise das variáveis, foi utilizada estatística descritiva. RESULTADOS: Realizaram-se mudanças e substituições de termos e expressões para obter equivalência cultural do WORC. Alteraram-se também os termos "não compreendidos" pelos pacientes de acordo com as sugestões feitas por eles. CONCLUSÃO: Após a tradução e adaptação cultural do questionário, foi concluída a versão em Português do WORC que está em processo de validação para ser utilizada no Brasil.

OBJECTIVE: To accomplish the translation of WORC (The Western Ontario Rotator Cuff Index) into Portuguese and its cultural adaptation, for future validation in Brazil. METHOD: The protocol applied consisted of: 1) Preparation; 2) Forward translation; 3) Back translation; 4) Cognitive Debriefing; and 5) Information report. After concluding the forward and back-translations, the versions were sent to the authors of the original questionnaire, who gave their approval for the study to be continued. The Portuguese version was applied to 35 patients with rotator cuff dysfunctions to verify the level of understanding of the instrument. The mean age of these patients was 57 years (SD = 13); 63 percent were female and 74 percent had not completed elementary school. The final Brazilian version of WORC was defined after achieving a "no comprehension" rate of less than 15 percent for each item. Descriptive statistics were used to analyze the variables. RESULTS: Some terms and expressions were changed or replaced to obtain cultural equivalence for WORC. The terms that were incomprehensible to the patients were changed in accordance with their own suggestions. CONCLUSION: After the translation and cultural adaptation of the questionnaire, the final Portuguese version of WORC was concluded and it is now undergoing validation for use in Brazil.

Risk adjusted surgical audit in gynaecological oncology: P-POSSUM does not predict outcome.

AIMS: To assess the Physiological and Operative Severity Score for the enumeration of mortality and morbidity (POSSUM) and its validity for use in gynaecological oncology surgery. METHODS: All patients undergoing gynaecological oncology surgery at the Northern Gynaecological Oncology Centre (NGOC) Gateshead, UK over a period of 12months (2002-2003) were assessed prospectively. Mortality and morbidity predictions using the Portsmouth modification of the POSSUM algorithm (P-POSSUM) were compared to the actual outcomes. Performance of the model was also evaluated using the Hosmer and Lemeshow Chi square statistic (testing the goodness of fit). RESULTS: During this period 468 patients were assessed. The P-POSSUM appeared to over predict mortality rates for our patients. It predicted a 7% mortality rate for our patients compared to an observed rate of 2% (35 predicted deaths in comparison to 10 observed deaths), a difference that was statistically significant (H&L chi(2)=542.9, d.f. 8, p<0.05). CONCLUSION: The P-POSSUM algorithm overestimates the risk of mortality for gynaecological oncology patients undergoing surgery. The P-POSSUM algorithm will require further adjustments prior to adoption for gynaecological cancer surgery as a risk adjusted surgical audit tool.

Unilateral groin and pelvic irradiation for unilaterally node-positive women with vulval carcinoma.

It is essential that any patient with resected vulval cancer and significant nodal disease receive optimal adjuvant treatment with radiation. Adequate radiotherapy for such patients with unilateral positive groin nodes has not been defined. Whether both groins and pelvic sidewalls should be irradiated or only the affected (node positive) side remains unclear. From our registry, we identified all patients with primary, previously untreated squamous cell carcinoma of the vulva undergoing bilateral inguinofemoral lymphadenectomy (superficial and deep nodes) and having unilaterally positive groin nodes treated with unilateral groin and pelvic radiotherapy (44 Gy in 22 fractions). Clinical and pathologic records were reviewed to identify the anatomical site and timing of recurrences in these patients and determine whether unilateral groin and pelvic irradiation was sufficient for disease control on the node-negative side. From 1983 to 2002, 20 patients with unilateral positive nodes treated with unilateral groin and pelvic irradiation were identified. Nineteen patients were classed as having FIGO stage III disease and one as FIGO stage IV due to involvement of the rectal mucosa. There were nine patients with disease recurrences in this group (45%). The disease-free interval ranged from 4 to 31 months (median time to recurrence, 9 months). All nine patients had local or regional failures, the most common site being the ipsilateral groin (six of nine patients). One patient was also found to have distant metastases. There were no recurrences noted in the contralateral (nonirradiated) groin or pelvic sidewall. Recurrence was generally fatal. Eight of the nine patients subsequently died of their disease. The ninth patient died of another cause. There was a high incidence of regional failure after unilateral groin and pelvic radiotherapy, but there were no recurrences on the nonirradiated, node-negative side. Although a small series, we speculate that there is no apparent disadvantage to administering unilateral adjuvant radiotherapy for unilaterally positive groin nodes and encourage further studies in order to more confidently determine whether the tendency observed in our center holds true.

Laparoscopically assisted radical vaginal hysterectomy vs. radical abdominal hysterectomy for cervical cancer: a match controlled study.

OBJECTIVES: The technical feasibility of laparoscopically assisted radical vaginal hysterectomy has been well described, but its advantages over the open technique remain largely unproven. We reviewed and compared our experiences with both approaches. METHODS: All patients undergoing laparoscopically assisted radical vaginal hysterectomy (LARVH) between 1996 and 2003 were identified and matched for age, FIGO stage, histological subtype and nodal metastases using a control group of women who underwent radical abdominal hysterectomy (RAH) during the same time period. RESULTS: Fifty-seven women were listed for LARVH, resulting in five conversions. Fifty cases were matched successfully using the criteria above. The majority of cases were FIGO stage 1B1. Statistically significant differences (P < 0.05) were present when the following were compared for LARVH vs. RAH: duration of surgery (median 180 vs. 120 min), blood loss (median 350 vs. 875 ml), hospital stay (median 5 days vs. 8 days) and duration of continuous bladder catheterisation (median 3 days vs. 7 days). There were no statistically significant differences with regard to nodal yield, completeness of surgical margins or perioperative complication rate. Four major complications (8%, three cystotomies and one enterotomy) occurred in the LARVH group and three in the RAH group (6%, one pulmonary embolism, one ureteric injury and one major haemorrhage). Three women in LARVH group had seen a specialist regarding postoperative bladder dysfunction, versus 12 in the RAH group (P = 0.04). No patients in the LARVH group reported constipation requiring regular laxatives, versus six in the RAH group (P = 0.03). Median follow-up was 52 months for LARVH and 49 months for RAH. There was no significant difference between recurrence rates or overall survival (94% for LARVH vs. 96% for RAH). CONCLUSIONS: Despite the inherent limitations of LARVH and its associated learning curve, the procedure conveys many advantages over the open technique in terms of blood loss, transfusion requirement and hospital stay. In addition, the incidence of postoperative bladder and bowel dysfunction appears low-suggesting improved quality of life-without compromising survival.

Drainage following radical hysterectomy and pelvic lymphadenectomy: dogma or need?

OBJECTIVE: To compare the incidence of lymphocyst formation and postoperative morbidity in patients drained or not drained following radical hysterectomy and pelvic lymph node dissection for cervical or endometrial malignancy. METHODS: A prospective study was undertaken of consecutive patients undergoing radical hysterectomy and pelvic lymphadenectomy at the Regional Department of Gynaecological Oncology, Gateshead, United Kingdom, between February 1992 and September 1994. A Piver type II procedure was performed with nonclosure of the vaginal cuff and pelvic peritoneum. Patients were randomized at the end of surgery to have either two suction drains inserted along the pelvic sidewalls or to have no drains inserted. The detection of lymphocysts was made by clinical examination and abdominal ultrasound scan performed 8 weeks postoperatively. RESULTS: Eight patients were excluded from the study when drains were deemed necessary to assess postoperative blood loss. Fifty-one were randomized to drains, and 49 to no drains. The detection of lymphocysts by ultrasound and clinical examination in the drained group (15.6 and 5.9%, respectively) was not significantly different from the group not drained (17.4 and 6.1%, respectively). There was no difference in postoperative morbidity in the two groups. CONCLUSION: There appears to be no advantage to the routine use of pelvic suction drainage following radical hysterectomy and pelvic lymphadenectomy.

"Microinvasive" adenocarcinoma of the cervix implanting in an episiotomy scar.

Episiotomy scar tumor implantation arising from a cervical carcinoma is rare. This report details a patient who developed an episiotomy implant from a "microinvasive" adenocarcinoma of the cervix 5 months following vaginal delivery. The implant behaved as a primary focus for metastases to the inguinal lymph nodes. Despite subsequent treatment with chemotherapy and radiotherapy the patient succumbed to the disease. The case is the first reporting a poor outcome following an episiotomy scar implant from a primary cervical adenocarcinoma. The poor outcome highlights the controversy regarding the use of the term microinvasive with respect to glandular cervical lesions.

An evaluation of the prognostic factors in metastatic gestational trophoblastic disease.

A retrospective evaluation of prognostic factors in 55 patients suffering from metastatic gestational trophoblastic disease (MGTD) treated by modified Bagshawe's CHAMOCA regimen was done. The prognostic significance of the eight prognostic factors in the WHO scoring system, number of sites of metastasis and FIGO staging were evaluated by univariate analysis using Chi-square test with Yates' correction and odds ratio and by multivariate analysis using Cox proportional hazard analysis and logistic regression analysis. In the univariate analysis, the intervals between antecedent pregnancy and the diagnosis of GTD, (P = 0.004) the level of hCG (P = 0.02) and the number of metastatic sites (P = 0.046) were significantly associated with death. In the multivariate analysis, only the interval between the antecedent pregnancy and the diagnosis and the level of hCG were significantly associated with death. Thus, it seems that the interval between antecedant pregnancy and the diagnosis and the level of hCG were the two most significant factors in predicting mortality in high risk MGTD. The WHO staging was more predictive of poor outcome than that of the FIGO staging in this group of patients.

Localization of sarcoma xenografts in nude mice with indium-111-labeled monoclonal antibodies.

A labeling method utilizing modified carbohydrate moieties in antibody heavy chains as radionucleotide binding sites was evaluated. Murine anti-sarcoma monoclonal antibody (MAb 19-24) was labeled with Indium-111 (111In) using this technique and subcutaneous human sarcoma xenografts were successfully localized in nude mice. A nonspecific monoclonal antibody BL-3 was used as a negative control. Tumor-to-blood ratios of radioactivity in the mice injected with 111In-labeled MAb 19-24 were significantly (P < 0.05) higher than those obtained with nonspecific MAb BL-3. Calculations of percentage injected dose of radioactivity per gram tissue showed relatively high specific uptake of MAb 19-24 in sarcoma xenografts. Radioactivity cleared from the blood rapidly and hepatic uptake of 111In-labeled antibodies was found to be relatively low. Biodistribution studies in normal mice with 111In-labeled antibodies showed only blood pool activity with no significant concentration of activity into organs. Therefore, immunoreactivity of the antibodies was retained after 111In-labeling utilizing this new technique, allowing specific binding of radiolabeled MAb to tumor xenografts with relatively low hepatic uptake.

Immunohistochemical and pharmacokinetic characterization of site-specific immunoconjugate 15A8-glycyl-tyrosyl-(N-epsilon-diethylenetriamine pentaacetic acid)-lysine derived from anti-breast carcinoma monoclonal antibody 15A8.

In this study, the breast carcinoma-reactive monoclonal antibody 15A8 and a site-specific immunoconjugate of the antibody, 15A8-glycyl-tyrosyl-(N-epsilon-diethylenetriamine pentaacetic acid)-lysine (15A8-GYK-DTPA), were characterized by immunohistological methods for reactivity with normal and neoplastic human tissues and normal cynomolgus monkey tissues. In addition, 15A8-GYK-DTPA labeled with 111In was assessed by in vivo imaging and pharmacokinetic studies for localization to human tumor xenografts in nude mice. The native antibody and the site-specific immunoconjugate exhibited similar limited reactivity with normal human tissues. Specifically, epithelial structures, including normal breast epithelium, lung alveoli, bronchial epithelium and glands, liver bile ducts, pancreatic ducts, kidney distal and collecting tubules, epidermal and esophageal epithelium, endometrial glands, and thymic Hassall's corpuscles, were reactive. Normal monkey tissues stained with 15A8 exhibited a similar pattern of reactivities. Antibody 15A8 reacted broadly with epithelium-derived tumors; more than 60% of the cells in all of the breast, colon, non-small cell lung, ovarian, prostate, bladder, and renal carcinomas tested expressed the antigen. In contrast, a variety of nonepithelial neoplasms, including lymphomas, melanomas, sarcomas, and small cell lung carcinomas, were nonreactive. 15A8-GYK-DTPA-111In administered i.v. rapidly localized to and imaged both MX-1 and MCF-7 human breast carcinoma xenografts in nude mice, reaching maximal levels of about 20% of injected dose/g of tumor within 4 days. No unusual localization to any nontumor tissue or organ was seen; the level of radioactivity in the normal tissues and organs was at or below that seen in the blood. Furthermore, the immunoconjugate did not accumulate in xenografts of the antigen-negative breast carcinoma ZR-75-1, which indicates that tumor localization was antigen specific. Pharmacokinetic studies in cynomolgus monkeys suggested that significant amounts of 15A8-GYK-DTPA-111In did not localize to normal epithelia and demonstrated that the immunoconjugate was not toxic. These findings suggest that antibody 15A8 may be useful in the diagnosis and therapy of breast cancer and possibly other carcinomas.

Immunohistochemical and pharmacokinetic characterization of the site-specific immunoconjugate CYT-356 derived from antiprostate monoclonal antibody 7E11-C5.

In this study, a site-specific immunoconjugate, designated CYT-356, of the prostate-reactive monoclonal antibody 7E11-C5 was characterized by immunohistological methods for reactivity with normal and neoplastic human tissues. In addition, CYT-356 labeled with 111In was assessed by in vivo imaging and pharmacokinetic studies for localization to human tumor xenografts in nude mice. The native antibody and the site-specific immunoconjugate exhibited similar patterns of reactivity with normal human tissues. Although the majority of tissues tested were negative, weak reactivity with cardiac muscle, proximal kidney tubules, and sweat glands was observed. Positive staining of normal prostate epithelial cells and glandular lumina and strong reactivity with a subset of skeletal muscle cells were also observed. CYT-356 reacted with 100% of prostate tumors examined but was negative on a variety of other neoplasms. Following i.v. administration, CYT-356-111In rapidly localized to and imaged LNCaP human prostate adenocarcinoma xenografts in nude mice, reaching maximal levels of about 30% of injected dose/g of tumor within 3 days. No unusual localization was seen to any nontumor tissue or organ; the level of radioactivity in the normal tissues and organs was at or below that seen in the blood. The localization to xenografts was antigen specific and the accessible binding sites in 100-200-mg tumors appeared to be saturated at an antibody dose between 10 and 100 micrograms. These findings suggest that the CYT-356 immunoconjugate may be useful in the diagnosis and therapy of prostate cancer.

Carbohydrate-derivatized immunoconjugate of the anti-(carcinoembryonic antigen) monoclonal antibody C46: immunohistological reactivity and pharmacokinetic comparison with a randomly derivatized C46 immunoconjugate.

In this study, a site-specific glycyl-tyrosyl-(N-epsilon-diethylenetriaminepentaacetic acid)-lysine (GYK-DTPA) immunoconjugate of the anti-carcinoembryonic antigen monoclonal antibody C46 (C46-GYK-DTPA) was characterized by immunohistological and immunofluorescence methods for reactivity with normal and neoplastic human tissues. In addition, pharmacokinetic studies assessed the ability of C46-GYK-DTPA labeled with 111In to localize to and image human tumor xenografts in nude mice. The native antibody and the site-specific immunoconjugate exhibited similar patterns of reactivity with normal human tissues. C46 did not bind to the surface of normal human granulocytes, which indicates lack of reactivity with normal cross-reacting antigen. C46-GYK-DTPA reacted with 100% of the colon, breast and renal carcinomas examined and with two of three lung carcinomas, but did not react with any sarcomas, melanomas or lymphomas examined. Intravenously administered C46-GYK-DTPA-111In rapidly localized to and imaged LS174T human colon adenocarcinoma xenografts in nude mice, reaching maximal levels of about 25% of injected dose/g tumor within 1 day. No unusual localization to any non-tumor tissue or organ was seen; the level of radioactivity in the normal tissues and organs was at or below that in the blood. The accessible binding sites in 1 g tumors appeared to be saturated at an antibody dose between 100 micrograms and 1000 micrograms/mouse. Further, in a direct in vivo comparison, the site-specific conjugate C46-GYK-DTPA had more favorable pharmacokinetics and better tumor localization than a randomly derivatized C46 immunoconjugate (C46-DTPA). These findings suggest that the site-specific immunoconjugate C46-GYK-DTPA may be useful in the diagnosis and therapy of colon cancer and other adenocarcinomas expressing carcinoembryonic antigen.

Radioimmunoscintigraphy and radioimmunotherapy in nude mouse models. Studies with site-specifically modified monoclonal antibodies.

Site-specific covalent modification of monoclonal antibodies at the oligosaccharide offers advantages over more conventional modification processes that involve direct attachment at tyrosine, lysine or glutamic/aspartic acid side chains. Using the site-specific modification process, attachment sites on the antibody are distal to the antigen-binding region. Thus, homogeneity of antigen-binding properties and affinity for the unmodified protein are preserved. Furthermore, higher derivatization ratios with no resultant loss of immunoreactivity can be achieved for monoclonal antibodies modified at the oligosaccharide. In vivo biodistribution and tumor localization studies in nude mouse models suggest that antibodies radiolabeled at their oligosaccharide might represent improved immunoscintigraphic reagents. In a variety of tumor xenograft models, site-specific modified 111In-labeled antibody conjugates localized to the tumor site with little non-specific localization in other tissues or organs. The degree of localization at the target site was substantially greater than that of 111In-labeled antibodies directly modified at the tyrosine side chain. Preliminary studies with 212Bi- and 90Y-labeled antibodies modified at the oligosaccharide indicate that both of these radioisotopes have immunotherapeutic potential. Because of its preferential uptake by the kidney, the use of 212Bi may be best suited for tumors localized within the peritoneal cavity, such as ovarian and colorectal carcinomas. The toxicity of 90Y at high specific activities suggests that a regimen of repeated smaller doses of this radioisotope is best suited for therapeutic use. Studies in tumor-bearing mouse models are currently underway to better define the optimal dosage and administration regimens for both of these radioisotopes when attached to site-specific modified antibodies.