Age‐ and site‐specific decline in insulin‐like growth factor‐I receptor expression is correlated with differential growth plate activity in the mouse hindlimb

The proximal and distal growth plates of the principal long bones do not contribute equally to longitudinal growth. Most forelimb elongation occurs at the shoulder and wrist, while most hindlimb growth occurs at the knee. This study examined whether insulin‐like growth factor‐I (IGF‐I), a potent growth regulator, could underlie this variation via differential receptor expression. The spatiotemporal distribution of the IGF‐I receptor (IGF‐IR) was mapped in hindlimb growth plates (overall and within regional zones) from immature mice using immunohistochemistry. Growth activity was assessed by size/morphology of the growth plate and proliferating cell nuclear antigen (PCNA) expression. Both IGF‐IR and PCNA staining declined considerably with age in the proximal femur and distal tibia (hip and ankle), but expression remained high in the more active distal femur and proximal tibia (knee) throughout growth. Growth plate size decreased with age in all sites, but the absolute and relative decline in IGF‐IR in the hips and ankles of older mice indicated a site‐specific loss of IGF‐I sensitivity in these less active regions. These results suggest that regulation of the IGF‐IR may at least partially mediate differential long bone growth, thereby providing a local mechanism for altering skeletal proportions absent modification of systemic hormone levels. Anat Rec, 2007. © 2007 Wiley‐Liss, Inc.     

噬菌体随机肽库分析HIV-1 p24抗原表位

目的：利用噬菌体随机肽库分析抗HIV－1核心区抗原p24单抗体在抗原上的识别位点。方法：用抗HIV－1 p24单抗2C7和3H10作为筛选分子，对噬菌体肽库进行生物洗（biopanning)，并通过DN测序、ELISA效价测定等对所获得的噬菌休克隆进行鉴定，最后对合成的7肽位点通过间接ELISA及免疫抑制试验进行血清学分析。结果：序列分析结果表明，单抗2C7和3H10在HIV－1 p24上的抗原识别表位的保守序列分别为DHPXPXX和XXXXKAF。分别合成这2个7肽氨基酸序列P－C1（DHPSPWG）和P－H3（SPWLKAFGGS），并分析其免疫学结合特性，结果表明与P－H3相比，单抗2C7的抗原识别表位P－C1的固相结合特性较好，固相P－C1检测血样，13份抗HIV阳性本中，12份为阳性（检出率为92．3％），19份抗HIV阴性样本中，仅1份为假阳性结果（特异性为94．7％），与P－C1相比，单抗3H10的抗原表位P－H3的固相结合能力极差，但液相结合活性较好，血样与P－H3的抑制试验表明，13份抗HIV阳性样本中12份样本对P－H3的抑制率大于60％（12／13），而9份抗HIV阴性样本中仅1份对P－H3的抑制率大于50％，结论：用抗HIV－1 p24单抗筛选噬菌体随机肽库，获得单抗在p24抗原上的识别表位的氨基酸序列，血清学结果表明这2个抗原表位存在于p24自然抗原上，在抗HIV－1的感染检测中具有潜在的应用价值。     

Antibody fragments selected by phage display against the nuclear localization signal of the HIV-1 Vpr protein inhibit nuclear import in permeabilized and intact cultured cells

The HIV-1 Vpr protein harbors a nuclear localization signal in its N-terminal domain. A peptide bearing this domain and which is designated VprN has been used as a target to screen a phage display single chain Fv (scFv) library. Here we report the isolation of anti-VprN scFv fragments from this library. The purified scFv fragments were able to bind the VprN peptide in an ELISA-based system and to inhibit VprN-mediated nuclear import in permeabilized as well as in intact microinjected cells. Furthermore, the anti-VprN scFv fragments recognized the full-length recombinant Vpr protein and inhibited its nuclear import. The same scFv fragments did not inhibit nuclear import mediated by the nuclear localization signal of the SV40 large T-antigen demonstrating a specific effect. The use of the described inhibitory anti-VprN scFv fragments to study nuclear import of viral karyophilic proteins and their therapeutic potential is discussed.     

特殊染色和免疫组织化学染色在肾上皮性肿瘤诊断中的应用

目的 探讨肾上皮性肿瘤的诊断及鉴别诊断。方法 对91例具有较完整病理资料的肾上皮性肿瘤进行了常规组织学观察,Mowy胶状铁染色及CD10、细胞角蛋白(CK)7、波形蛋白免疫组织化学染色。结果 91例肾上皮性肿瘤中肾透明细胞癌78例,占86％;肾乳头状癌8例,占9％;肾嫌色细胞癌4例,占4％;肾嗜酸细胞腺瘤1例,占1％。78例肾透明细胞癌中CD10、波形蛋白阳性分别为63例(81％)和69例(88％),主要表现为细胞膜阳性。74／78肾透明细胞癌CK7呈阴性,17／17 Mowy胶状铁染色阴性或呈灶状粗颗粒。肾嫌色细胞癌4例,CK7均呈胞膜阳性;Mowy胶状铁染色表现为胞质内蓝色细网状;4例CD10肿瘤细胞膜均阴性,波形蛋白阴性。肾嗜酸细胞腺癌1例CD10,CK7,波形蛋白,Mowy胶状铁染色均为阴性或不特异。结论 在常规诊断肾上皮性肿瘤有困难时免疫组织化学CD10,CK7,波形蛋白染色以及Mowy胶状铁染色对鉴别诊断有帮助。     

蝶窦垂体腺瘤的临床病理学观察

目的 探讨蝶窦垂体腺瘤(sphenoid sinus pituitary adenoma,SSPA)的临床病理特征,提高对蝶窦垂体腺瘤的认识和病理诊断水平.方法 对12例SSPA进行临床、组织病理学和SP法免疫组化染色(PRL、hGH、ACTH、LH、TSH、FSH等).结果 12例SSPA,男5例,女7例,年龄39～66岁,平均48.4岁,中位年龄55.5岁.其中原发于蝶窦和原发于垂体窝侵至蝶窦者各6例,原发于蝶窦者年龄较原发于垂体窝者偏大.临床症状多样.镜下,原发于蝶窦的垂体腺瘤的形态学与蝶鞍垂体腺瘤相同;SSPA可伴有较明显的慢性炎症细胞浸润(2/12,16.67%).10例行免疫组织化学染色的结果,瘤细胞PRL阳性6例;瘤细胞各项标记物均阴性2例;瘤细胞ACTH阳性、hGH阳性各1例.结论原发于蝶窦的垂体腺瘤比较少见,主要根据光镜及免疫组化确诊,应注意与慢性蝶窦炎和浆细胞瘤鉴别.     

Rapid accurate absolute granulocyte count determination during severe leukopenia using the flow cytometer

The absolute granulocyte count (AGC) in 125 blood samples from patients with total white blood cell counts of less than 1,000/microliter was estimated using three different methods, which were then compared for efficiency and accuracy. The three methods were 25 cell differential counts using Wright's-stained blood smears, granulocyte percentage estimates from WBC counting chambers, and combined narrow- and wide-angle light-scatter characteristics determined on a flow cytometer. A survey of clinical laboratories at University Hospital Cancer Centers revealed that the smear differential was the most-often-used method in those laboratories even when less than 25 cells could be counted. Consequently the data obtained from the counting chamber and flow cytometer methods were compared to the smear differential "standard" using linear regression, and outliers were identified. There was good correlation between AGC determined by smear differential and WBC counting chamber (correlation coefficient .911) and excellent correlation between the AGC determined by smear differential and the flow cytometer method (correlation coefficient .970). The flow cytometer method used in this investigation required minimal specimen preparation, and test results were available at a rate of 60 seconds/sample. The ease of sample preparation, speed, and statistical reliability of test results makes the flow cytometer an attractive alternate method of determining granulocyte counts on leukopenic patients as compared to the stained blood smear differential.     

Histological characterization of neurones innervating functionally different muscles of Locusta

The innervation of four functionally different muscles (subalar, remotor 1, remotor 2, pleuroalar), all served by the same nerve branch, was studied in both winged segments of the locust, Locusta migratoria. Several anatomical techniques were applied: With the cobalt backfill and silver intensification technique four cell types (motoneurone, dorsal unpaired median neurone, common inhibitory neurone, and small median neurone) were demonstrated. Serial sections enabled the morphology of the motoneurones to be described in more detail and in respect to a possible functional organization of the arborizations. A differential staining technique allowed us to stain various neurones in different colours in the same preparation. With this technique the anatomy of both the "rostral" and the "caudal" subalar motoneurones could be described in parallel, thus avoiding errors in comparison due to possible individual variations from preparation to preparation. Axon counts in the peripheral nerve branch enabled us to compile a list of the total innervation for each muscle. Results from other orthopterans are integrated and whether differences in the dendritic fields might be of functional significance is discussed.     

Macromolecular DNA-damage in murine and human leukemic and lymphoid cells after in vitro exposure to ASTA Z 7557 (INN mafosfamide)

Summary Cyclophosphamide (CPA) is widely used against leukemic and lymphoproliferative diseases, but in vitro studies on response to this agent so far have been limited to instable derivatives with poor galenic properties. ASTA Z 7557 is a newly synthesized activated cyclophosphamide that circumvents the need for hepatic activation and has good stability. The critical cytotoxic lesions after exposure to bifunctional alkylating agents presumably are DNA interstrand crosslinks (ISC). We have, therefore, examined the formation and apparent removal of ISC after in vitro treatment with ASTA Z 7557 by use of the highly sensitive alkaline elution technique. Survival of murine L1210 cells was determined after 1 hour in vitro exposure with a D 37 value of 5.7 g/ml (from the initial shoulder part of the survival curve) and a Do value of 1.5 g/ml (from the exponential part of the curve). Previous labelling of L1210 cells by ¹²⁵IUdR simplified the alkaline elution procedure but there was some cytotoxicity of the radiochemical itself with a reduction of cloning efficiency from 77% to 61 %. The maximum of ISC was observed at 6 h after initiation of treatment with much of the damage apparently removed at 24 h. The simultaneous presence of DNA single strand breaks (SSB), however, confounds the analysis of DNA damage at 24 h and early cytolysis and unaided death of human lymphocytes often preclude the analysis of macromolecular damage at this time. Human peripheral blood cells isolated from patients with leukemic or lymphoproliferative diseases showed a remarkable heterogeneity with regard to the formation of ISC at 3 h. Thus, analysis of macromolecular damage may become an additional prognostic factor for response to CPA beyond the morphologic classification of these diseases.     

Are video display terminals a source of increased PCB concentration in the working atmosphere?

Summary In response to the question: Are datascreen terminals a source of increased PCB concentrations in the working atmosphere? a study of PCB emissions from video display terminals (VDT) was undertaken. Emissions of 2.4 to 8.1 ng PCB/h were observed from VDT located in a building (1) where the mean PCB level in the air was 46 ng PCB/m³ during the test period, whereas no PCB emissions were detected from VDT located in a building (2) where no PCB could be detected in the ambient air. However, both the air and the VDT from building 2 were found to be contaminated with polycyclic aromatic hydrocarbons. We conclude that the observed PCB emissions from VDT are the result of the vapourization of PCB deposited onto the VDT from the PCB contaminated air and do not originate from the electrical components of the VDT.     

An Epidemiology Model for Estimating the Numbers of US Patients With Multiple Myeloma by Line of Therapy and Treatment Exposure

ObjectivesEstimates on the distribution of patients with multiple myeloma (MM) by line of therapy (LOT) are scarce and get outdated quickly as new treatments become available. The objective of this study was to estimate the number of patients with MM by LOT and the number of patients who have received at least 4 previous LOTs including proteasome inhibitors, immunomodulatory agents, and anti-CD38 monoclonal antibodies (mAbs).MethodsA compartmental model was developed to calculate the number of patients by LOT. Two pathways were considered based on stem cell transplant eligibility, and at each pathway, treatments were stratified in 2 types: anti-CD38 mAbs or other. The model population was stratified into 4 subgroups based on age and cytogenetic risk. Model inputs were informed from real-world evidence.ResultsThe model estimated that, in 2020, 126 869 patients were living with MM in the United States. Of these, 105 701 received treatment in any LOT, with 56 959, 27 252, 11 258, and 5217 in lines 1 to 4, respectively, and 5015 in line 5 or beyond. The model estimated that 3497 patients received at least 4 previous LOTs including proteasome inhibitors, immunomodulatory agents, and anti-CD38 mAbs. The model overall prevalence predictions aligned well with publicly available estimates.ConclusionsThis study proposes a novel framework to estimate MM prevalence. It can assist clinicians to understand future trends in MM epidemiology, healthcare systems to plan for future resource use allocation, and payers to quantify the budget impact of new treatments.