Monoclonal antibodies against transferrin. Precipitating mixtures and lack of inter-species cross-reactivity

Five stable hybridoma lines were prepared using the myeloma cell line P3-X63-Ag.653 and spleen cells of mice hyperimmunized by pig transferrin. All hybridomas grew well in mouse peritoneal cavity and produced antibodies of the IgG₁ subclass. Antibody preparations obtained from ascitic fluids tested for their capacity of antigen precipitation. No precipitation was obtained with single antibodies and with pairs of antibodies. Three out of 10 possible triads gave clear and sharp precipitation zones and rings in immunodiffusion tests performed in agar gel. All 5 antibodies were shown by quantitative enzyme-immunoassay to be specific for pig transferrin: no cross-reaction was obtained with mouse, human, horse and sheep transferrins.     

Expression of P fimbriae of uropathogenic Escherichia coli strains

The occurrence of P fimbriae in a total of 222 uropathogenic Escherichia coli (UPEC) strains was investigated. Out of the total, 31 (14%) were P fimbriated. Of 24 pyelonephritogenic E. coli strains, three (13%) with P fimbriae occurred in children with clinical pyelonephritis, and of 198 E. coli strains 29 (15%) occurred in children with cystitis. Prevalence of P fimbriae of E. coli strains was found to be quite similar in patients with cystitis and pyelonephritis     

Revival of the natural cycles in in-vitro fertilization with the use of a new gonadotrophin-releasing hormone antagonist (Cetrorelix): a pilot study with minimal stimulation

Natural cycles were abandoned in in-vitro fertilization (IVF) embryo transfer, due to premature luteinizing hormone (LH) surges--and subsequent high cancellation rates. In this study, we investigated the administration of a new gonadotrophin-releasing hormone antagonist (Cetrorelix) in the late follicular phase of natural cycles in patients undergoing IVF and intracytoplasmic sperm injection (ICSI). A total of 44 cycles from 33 healthy women [mean age 34.1 +/- 1.4 (range 26-36) years] were monitored, starting on day 8 by daily ultrasound and measurement of serum concentrations of oestradiol, LH, follicle stimulating hormone (FSH) and progesterone. When plasma oestradiol concentrations reached 100-150 pg/ml, with a lead follicle between 12-14 mm diameter, a single injection (s.c.) of 0.5 mg (19 cycles) or 1 mg (25 cycles) Cetrorelix was administered. Human menopausal gonadotrophin (HMG; 150 IU) was administered daily at the time of the first injection of Cetrorelix, and repeated thereafter until human chorionic gonadotrophin (HCG) administration. Four out of 44 cycles were cancelled (9.0%). No decline in follicular growth or oestradiol secretion was observed after Cetrorelix administration. A total of 40 oocyte retrievals leading to 22 transfers (55%) was performed. In 10 cycles (25%), no oocyte was obtained. Fertilization failure despite ICSI occurred in six cycles (15%). In two patients the embryo was arrested at the 2 pronuclear (PN) stage. The stimulation was minimal (4.7 +/- 1.4 HMG ampoules). A total of seven clinical pregnancies was obtained (32.0% per transfer, 17.5% per retrieval), of which five are ongoing. Thus, a spontaneous cycle and the GnRH antagonist Cetrorelix in single dose administration could represent a first-choice IVF treatment with none of the complications and risks of current controlled ovarian hyperstimulation protocols, and an acceptable success rate.     

Adult pancreas generates multipotent stem cells and pancreatic and nonpancreatic progeny

Strategies designed to produce functional cells from stem cells or from mature cells hold great promise for treatment of different cell-degenerative diseases. Type 1 and type 2 diabetes are examples of such diseases. Although different in origin, both involve inadequate cell mass of insulin-producing beta cells, the most abundant cell type of pancreatic islets of Langerhans. Practical realization of such strategies is highly dependent on the elucidation of physiological mechanisms responsible for generation of new beta cells in the pancreas, which at this time are poorly defined. The in vitro differentiation systems allowing generation of new beta cells provide a valuable experimental tool for studying these mechanisms. Few such systems are currently available. In this work, we present an in vitro differentiation system, derived from adult mouse pancreas, capable of generating insulin-producing beta-like cells, which self-organize into islet-like cell clusters (ILCCs) during the course of the culture. Surprisingly, we found that along with the ILCCs, multiple cell types with phenotypic characteristics of embryonic central nervous system and neural crest are also generated. Moreover, several embryonic stem cell-specific genes are induced during the course of these cultures. These results suggest that the adult pancreas may contain cells competent to give rise to new endocrine and neural cells.     

Grandparental genotyping enhances exome variant interpretation

Trio exome sequencing is a powerful tool in the molecular investigation of monogenic disorders and provides an incremental diagnostic yield over proband‐only sequencing, mainly due to the rapid identification of de novo disease‐causing variants. However, heterozygous variants inherited from unaffected parents may be inadvertently dismissed, although multiple explanations are available for such scenarios including mosaicism in the parent, incomplete penetrance, imprinting, or skewed X‐inactivation. We report three probands, in which a pathogenic or likely pathogenic variant was identified upon exome sequencing, yet was inherited from an unaffected parent. Segregation of the variants (in NOTCH1, PHF6, and SOX10) in the grandparent generation revealed that the variant was de novo in each case. Additionally, one proband had skewed X‐inactivation. We discuss the possible genetic mechanism in each case, and urge caution in data interpretation of exome sequencing data. We illustrate the utility of expanding segregation studies to the grandparent generation and demonstrate the impact on exome interpretation strategies, by showing that objective genotype data can overcome subjective parental report of lack of symptoms.     

中药四性的生物热动力学研究（Ⅱ）参叶和参花药性的微量量热学比较

目的 从生物物时化学的角度,阐明中药四性（寒、热、温、凉）的客观真实性。方法 利用微量量热法,测定了大肠杆菌在不同条件下的生长热谱曲线,得到了相应的生物热动力学参数和生长速率常数（K）、抑制率（I）、半抑制率（IC50）、热焓（△H）。根据生物热动力学通用数值,结合本草文献报道,综合分析了参叶和参花药性的差异所在。结果 参叶和参花抑制细菌生长的热谱曲线,其形状基本相同,随着药物浓度的加大,代谢过程的停滞期延长,生长代谢峰后移;其移力学参数势焓（△H）存在较稳定的差异--参叶降低了细菌生长代谢过程中热量的释放,而参花增加了细菌生长代谢过程中热量的释放。结论 微量量热法可作为刻划中药药性的一个新的有效手段,亦可作为中药活性筛选的新工具。     

Vascularisation des glandes surrénales

Résumé La glande surrénale de l'homme réunit deux structures initialement sans rapport physiologique. Cette dualité phyllogénique et embryologique se retrouve dans sa structure vasculaire de façon paradoxale. Les artères métamériques de l'organe phéochrome irriguent en priorité le cortex. L'axe veineux de l'ébauche mésenchymateuse draine le réseau des capillaires sinusoïdaux vers la médulla. Le réseau lymphatique est quasi inexistant au niveau de la médulla. Bien développé dans le cortex, il se draine presque exclusivement de façon centrifuge le long des axes artériels. A côté de son intérêt anatomo-chirurgical évident, l'étude de la vascularisation des glandes surrénales se doit de répondre aux besoins des investigations morphologiques vasculaires. La connaissance des variations artérielles et de leur fréquence, des territoires glandulaires de distribution, des voies anastomotiques de suppléance et des regroupements typologiques a retenu notre attention. Elle facilitera la lecture des artériographies globales toujours difficile dans le plan rétro-péritonéal. Elle guidera l'angiographie artérielle sélective. La phlébographie cave occlusive permet d'obtenir des opacifications remarquables des veines centrales, en particulier à gauche. Cette méthode est utile dans l'étude des tumeurs rétro-péritonéales et dans l'exploration des masses ganglionnaires lombo-aortiques susceptibles d'imprimer leur empreinte sur les structures veineuses. Elle a d'autant plus d'intérêt que la lymphographie pédieuse qui opacifie bien les chaînes latéro-aortiques gauche et latérocave à droite, ne permet pas, sauf reflux à contrecourant, une étude de l'important carrefour inter-aortico-cave sous-rénal. Des cathétérismes et des prélèvements électifs au niveau des différentes voies excrétrices de cette glande endocrine (veines et lymphatiques) peuvent aider à une meilleure connaissance de la physiologie et de la pathologie de l'organe. Les voies de dérivations veineuses des glandes surrénales ont été étudiées dans cet esprit.     

Prospective Evaluation of Prostate Specific Antigen (PSA), PSA Density,Free-to-Total PSA Ratio and a New Formula (Prostate Malignancy Index) for Detecting Prostate Cancer and Preventing Negative Biopsies in Patients with Normal Rectal Examinations and Intermediate PSA Levels

Objective: To improve the specificity and sensitivity of prostatic cancer detection, we prospectively evaluated total prostate specific antigen (PSA) level, PSA density, free-to-total PSA ratio and a new formula called prostate malignancy index (PMI) as a discriminator of prostate cancer in patients with intermediate PSA levels and normal digital rectal examinations. Materials and methods: Between November 1995 and October 1997, 95 patients who had serum PSA levels of 4.0 to 10.0 ng/ml with normal digital rectal examinations were prospectively evaluated. All patients underwent one or two times transrectal ultrasound guided prostate biopsies. Based on age specific reference range of PSA, PSA density and % free PSA ratio, PMI was calculated for each patient. The free and total serum PSA concentrations were determined by an Immulite assay system. (Diagnostic Product Corp., Los Angeles, California). Results: Overall 20 of 95 (21%) patients had prostate cancer. There were no significant differences in patient mean age and mean total PSA between those with benign and those with malignant biopsies (p>0.05). However, there were significant differences in mean PSAD, mean free-to-total PSA ratio and mean PMI (p<0.01, p<0.05, p<0.01, respectively). Benign condition specificities for PM index, percent free PSA, PSA density and total PSA at a 90% sensitivity for prostate cancer were 48%, 10.6%, 8% and 4%, respectively. Of 95 patients, 27 (28.4%) had a PMI of equal or more than 3.1, including 12 of 75 (16%) with negative biopsy and 15 of 20 (75%) with positive biopsy. Furthermore a cutoff MI 0.86 P correctly identified 24% of benign cases without missing any prostate cancer cases. The comparison of receiver operating characteristic (ROC) curve areas showed that PMI was better than total PSA (p<0.01). Although, the area under the ROC curve of % free PSA and PSAD were higher than the area of total PSA, these differences were not statistically significant (p>0.05). Conclusions: We concluded that the prostate malignancy index could be utilized to differentiate benign conditions from prostate cancer in patients with intermediate PSA levels and normal digital rectal examination. Also significant numbers of negative biopsies can be prevented in these patients.     

Mouse mammary tumor virus-like gene sequences in breast tumors of Australian and Vietnamese women.

PURPOSE: There is considerable evidence that the presence of mouse mammary tumor virus (MMTV)-like gene sequences in human breast cancer is highly associated with human breast carcinoma. Previous studies have found MMTV-like gene sequences in 38% of breast cancer tissue from United States women. The prevalence of these sequences in Australian and Vietnamese women has never been reported. EXPERIMENTAL DESIGN: Using PCR and primers that amplify MMTV-like gene sequences, we tested cancerous and benign breast tissue from Caucasian-Australian, Vietnamese-Australian, and Vietnamese women. RESULTS: MMTV-like gene sequences were amplified in 19 of 45 (42.2%) archival breast cancer biopsy tissues from Caucasian-Australian women, but only 1 of 120 (0.8%) and 0 of 41 breast cancer biopsy tissues from Vietnamese and Vietnamese-Australian women, respectively. The same sequences were found in only 2 of 111 (1.8%) and 0 of 60 normal (benign) breast tissue samples from Australian and Vietnamese women, respectively. CONCLUSIONS: MMTV-like gene sequences are found in only some human populations and are rarely found in normal human breast tissue from all populations, suggesting they are not present in the normal human genome and have been acquired.