[Recurrent intracerebral haemorrhage in a 24-year-old female patient]. / Rezidivierende intrazerebrale Blutungen bei einer 24-jährigen Patientin.

A 24-year-old female patient from Sierra Leone was referred to the authors' hospital after several unclear intracerebral bleeding events and an echogenic structure on the aortic valve. The patient was receiving oral anticoagulation therapy due to paroxysmal atrial fibrillation and left ventricular noncompaction. Fluorescence in situ hybridization in combination with polymerase chain reaction and sequencing revealed infective endocarditis of the mitral and aortic valve caused by Bartonella quintana. In retrospect, the intracerebral bleeding events could be identified as septic emboli with secondary haemorrhagic transformation under anticoagulation therapy. The patient showed significant clinical improvement and no further bleeding events occurred after receiving biological mitral and aortic valve replacement and several weeks of doxycycline and gentamicin antibiotic therapy.

A novel BCMA/CD3 bispecific T-cell engager for the treatment of multiple myeloma induces selective lysis in vitro and in vivo.

This corrects the article DOI: 10.1038/leu.2016.388.

A novel BCMA/CD3 bispecific T-cell engager for the treatment of multiple myeloma induces selective lysis in vitro and in vivo.

B-cell maturation antigen (BCMA) is a highly plasma cell-selective protein that is expressed on malignant plasma cells of multiple myeloma (MM) patients and therefore is an ideal target for T-cell redirecting therapies. We developed a bispecific T-cell engager (BiTE) targeting BCMA and CD3ɛ (BI 836909) and studied its therapeutic impacts on MM. BI 836909 induced selective lysis of BCMA-positive MM cells, activation of T cells, release of cytokines and T-cell proliferation; whereas BCMA-negative cells were not affected. Activity of BI 836909 was not influenced by the presence of bone marrow stromal cells, soluble BCMA or a proliferation-inducing ligand (APRIL). In ex vivo assays, BI 836909 induced potent autologous MM cell lysis in both, newly diagnosed and relapsed/refractory patient samples. In mouse xenograft studies, BI 836909 induced tumor cell depletion in a subcutaneous NCI-H929 xenograft model and prolonged survival in an orthotopic L-363 xenograft model. In a cynomolgus monkey study, administration of BI 836909 led to depletion of BCMA-positive plasma cells in the bone marrow. Taken together, these results show that BI 836909 is a highly potent and efficacious approach to selectively deplete BCMA-positive MM cells and represents a novel immunotherapeutic for the treatment of MM.

UPA and PAI-1 analysis from fixed tissues - new perspectives for a known set of predictive markers.

The urokinase-type plasminogen activator (uPA) and its main inhibitor PAI-1 play key roles in tumor-associated processes such as the degradation of the extracellular matrix (ECM), tissue remodeling, cell adhesion and migration. Elevated expression of both molecules is known to correlate with negative outcomes in node negative breast cancer. To date, these molecules are the only prognostic markers to have reached the highest level of evidence (LOE I) in multi-centered clinical trials for prognosis of node negative breast cancer. Unfortunately, the clinical utility of these molecules as markers is limited by the use of enzyme-linked immunoassay (ELISA) tests for their detection. The ELISA relies on the use of fresh or frozen tissue, which are rarely available in routine clinical settings. In this review article, we provide an overview of the clinical relevance of uPA and PAI-1 and present alternative methods for their detection. Common uPA and PAI-1 detection methods discussed in literature include RT-PCR-based assays and classical immunohistochemistry approaches. In recent years, attempts have been made to isolate and analyze proteins of formalin fixed, paraffin embedded (FFPE) tissues. These new methods are of special interest because up to now neither RT-PCR nor immunohistochemistry are recommended for the detection of uPA and PAI-1. Here, we present an approach for the analysis of uPA and PAI-1 directly from FFPE tissues that may eventually overcome the limitations of current assays and make the use of both markers widely available for routine prognosis and therapy decisions for breast cancer patients.

[Intraoperative volumetry in ENT surgery. Objectivation of surgical success by the volume control system]. / Intraoperative Volumetrie in der HNO-Chirurgie. Objektivierung des Operationserfolgs mit Volume-Control-System.

PROBLEM: At present no procedure exists to measure distances or volumes in endoscopic or otherwise limited surgical approaches directly and with high accuracy. Here a laser measuring system is evaluated for the first time as a clinical application in ENT surgery. MATERIAL AND METHODS: The volume control system (VCS) measures with the help of automatic recognition of laser measuring points in the surgical situs. A lab test examines the accuracy and the precision at anatomically accurate paranasal sinus and tympanic cavity models under flexible endoscopic visualization. The true values are known in each case as calibrated distances. Thus 90 values were available for evaluation. The clinical trial serves as proof of the intraoperative applicability and includes 32 patients. RESULTS: The measurements in the lab test resulted in an average deviation from the true value at a maximum of 7.1%. The precision was between 0.2 and 0.5 mm. In the clinical setting the system could be used in all 32 patients. Altogether 97 measured values could be included. The VCS functioned without system failures. The additional time required for setting up amounted to less than 2 min. The manageability of the flexible endoscope was reduced because of the length and the difficulty in controlling the adjustment. The additional intraoperative time required for collection of the measured values was less than 4 min in each case. Many results led to clinically relevant interpretations with intraoperative consequences. DISCUSSION: The VCS shows for the first time an intraoperatively applicable measuring function for distances, surfaces and volumes. There is a multiplicity of meaningful applications in ENT and in other surgical disciplines. The available study has proved the concept.

Quantitative protein analysis from formalin-fixed tissues: implications for translational clinical research and nanoscale molecular diagnosis.

Owing to its cross-linking effects, it is currently believed that formalin fixation of routinely processed tissues in the clinic prevents protein extraction and profiling. The aim of our study was to develop a robust, fast, standardized, and easy to use technique for the solubilization of non-degraded, full length, and immunoreactive proteins from formalin-fixed tissues for western blot and protein microarray analysis. Sections of routinely processed formalin-fixed and paraffin-embedded tissues of various origin were analysed. After deparaffination, tissues were manually dissected from the slides and transferred into an optimized protein extraction buffer system. Proteins were solubilized and subsequently analysed by western blot and reverse phase protein microarrays. We succeeded in isolating non-degraded, soluble, and immunoreactive proteins from routinely processed formalin-fixed tissues. We were able to detect membrane, cytoplasmic and nuclear proteins at the expected molecular weight. No differences were found in the protein yield and protein abundances between fresh frozen and formalin-fixed tissues. Using western blots and reverse phase protein microarrays, the receptor tyrosine kinase HER2, an important protein target for antibody based cancer treatment, was reliably measured in formalin-fixed breast cancer biopsy samples when compared with measurement by immunohistochemistry and fluorescence in situ hybridization; remarkably, immunohistochemically equivocal cases (score 2+) can be categorized according to HER2 protein abundance. Our new clinically orientated multiplexed protein measurement system may be generally applicable to determine the relative abundances of known disease-related proteins in small amounts of routinely processed formalin-fixed tissue samples for research and diagnosis. This technique may also be used to identify, characterize, and validate known and new protein markers in a variety of human diseases.

Clinical proteomics: new trends for protein microarrays.

Protein microarrays are an emerging class of nanotechnology for tracking many different proteins simultaneously. Much progress has been made for applications in basic sciences. Translation of these methods for the treatment of patients, however, is slow, because the realities in the clinic are rarely taken into account, and proteomic changes in cultured cell lines might not fully reflect human diseases due to the lack of the tissue microenvironment. In this review, we summarise current protein microarray approaches that are being developed for profiling tissues and histopathologically defined cell populations from cancer patients. We provide an overview of clinical applications for protein lysate microarrays and discuss the power of this technology for the discovery of disease markers for cancer diagnosis and individualised treatment.

Cell cycle progression of chronic lymphocytic leukemia cells is controlled by cyclin D2, cyclin D3, cyclin-dependent kinase (cdk) 4 and the cdk inhibitor p27.

B-CLL cells are arrested in G0/early G1 phase of the cell cycle and are characterized by a marked hyporesponsiveness towards a variety of polyclonal B cell activators. We have previously demonstrated that costimulation with CpG-ODN and IL-2 can overcome this proliferative defect. Cyclin D3 is the principal D-type cyclin which mediates G1 progression in normal B cells, but in B-CLL cells both cyclin D2 and cyclin D3, were strongly upregulated upon stimulation. Both cyclins were associated with cdk4 but not with cdk6, which is the catalytic partner of D-type cyclins in normal B cells. Moreover, immune complexes consisting of cyclin D2 and cdk4 or cyclin D3 and cdk4 were both functional and phosphorylated the RB protein in vitro. The cell cycle inhibitor p27 plays a pivotal role in cell cycle progression of B lymphocytes and has been shown to be overexpressed in B-CLL cells. P27 was rapidly downregulated in B-CLL cells even when stimulated with a non-CpG-ODN or IL-2 alone, while only moderate regulation could be observed in normal B cells. Taken together, our findings demonstrate that regulation of early cell cycle progression differs between B-CLL cells and normal B cells. These findings do not only contribute to the understanding of B-CLL pathophysiology, but might ultimately lead to the identification of new therapeutic targets.

Intra-tracheal delivery strategy of gentamicin with partial liquid ventilation.

Patients with pulmonary infection often present with ventilation and perfusion abnormalities, which can impair intravenous antibiotic therapy. Intra-tracheal (i.t.) administration has met with obstacles, such as inadequate delivery to affected lung regions and the disruption of gas exchange. We hypothesized that i.t. administration of a gentamicin (G)/perfluorochemical (PFC) suspension (G/PFC) would effectively deliver and distribute gentamicin to the lung, while maintaining gas exchange and non-toxic serum levels. In addition, we sought to compare serum G and lung levels and distribution of G when G/PFC is administered at the initiation of partial liquid ventilation (PLV) vs. during PLV. To test this hypothesis, 17 newborn lambs were ventilated by PLV with perflubron (LiquiVent) for 4 h using three different G (5 mg kg-1) administration techniques: i.t. slow-fill (SF) (n = 6; G/PFC over 15 min at start of PLV), i.t. top-fill (TF) (n = 6; G/PFC 10-65 min after start of PLV), intravenous (i.v.) (n = 5, aqueous injection at start of PLV). Serum levels of gentamicin were obtained 1, 15, 30 and 60 min after administration, and hourly there after for the remainder of the protocol (4 h). Arterial blood gas and pulmonary function measurements were obtained throughout the protocol. At the conclusion of the protocol, representative samples from each lung lobe, the brain and kidney were homogenized and assayed for gentamicin. All results are presented as the mean +/- SEM; P < 0.05. Over time, serum gentamicin levels were greatest (P < 0.05) in i.v. (11.0 +/- 2.3 micrograms ml-1), followed by TF (2.3 +/- 0.1 micrograms ml-1) and SF (0.8 +/- 0.1 microgram ml-1). The percentage of the administered dose remaining in the lungs after 4 h was greater (P < 0.05) following i.t. delivery (SF 23.8 +/- 4.3%, TF 13.7 +/- 2.5%) as compared to i.v. (3.7 +/- 0.5%). These findings suggest that for a given dose of G, both SF and TF delivery methods of G/PFC can enhance pulmonary, relative to systemic, antibiotic coverage.

Assessment of a geographically targeted field intervention on gonorrhea incidence in two New York State counties.

BACKGROUND AND OBJECTIVES: Using study findings that demonstrated the importance of core group members in gonorrhea transmission, in 1984, one New York State county changed its approach toward gonorrhea control by targeting its field intervention activities toward infected persons from a geographic core area. In late 1988, New York State experienced an epidemic increase in the number of syphilis cases. In response, the New York State sexually transmitted disease (STD) control program enacted a Syphilis Initiative, which required the diversion of field staff from gonorrhea to syphilis control activities for a 3-year period. Each of these events held the possibility of impacting gonorrhea incidence in this county. GOAL OF THIS STUDY: To evaluate the impact of core interventions on reducing gonorrhea incidence as compared to traditional nontargeted field intervention methods and to determine the influence on gonorrhea incidence of diverting field activities from gonorrhea to syphilis case finding. STUDY DESIGN: A Poisson regression method was used to estimate gonorrhea incidence for a 22-year period in two similar counties: one county that used core intervention and one that applied traditional case-finding methods. The impact of core intervention was estimated in terms of the reduction in the gonorrhea incidence rate from the preintervention incidence rates. RESULTS: After initiation of the core intervention, the relative risk of gonorrhea decreased by 61%. Between 30 % to 40% of the total reported cases were interviewed for contacts annually during the intervention period. In the control county, the relative risk was reduced by 50% despite a significantly higher percentage of annual case interviews (60%-70%). In addition, a small change in the definition of core (from census tracts encompassing 50% of gonorrhea cases to 30%-35%) during the Syphilis Initiative led to a significant increase (16%) in the relative risk of gonorrhea. CONCLUSION: Targeting partner notification activities toward a geographic core area population appears effective in reducing the risk of gonorrhea, and it was more efficient because the overall percentage of cases interviewed was smaller than in a county using a nontargeted approach. Diversion of staff during a syphilis epidemic, combined with a narrowing of the geographic scope of the core intervention, was associated with an increase in gonorrhea incidence.

Differential cholera-toxin sensitivity of supraspinal antinociception induced by the cannabinoid agonists delta9-THC, WIN 55,212-2 and anandamide in mice.

Intracerebroventricular (i.c.v.) administration to mice of delta9-tetrahydrocannabinol (delta9-THC), WIN 55,212-2 or the endogenous cannabinoid anandamide induced dose-related antinociception in the 55 degrees C warm-water tail-flick test. Pretreatment (24 h, i.c.v.) with pertussis toxin dose-dependently reduced the antinociceptive effect of delta9-THC (955 nmol), WIN 55,212-2 (30 nmol) and anandamide (135 nmol) (IC50 = 0.13, 5.5, and 0.32 nmol, respectively). In contrast, pretreatment (24 h, i.c.v.) with cholera toxin (0.1-3.0 mg) reduced the antinociception of WIN 55,212-2, had minimal effect on delta9-THC, and dose-dependently increased the antinociception of anandamide (ED50 = 0.50 nmol). These data suggest differences in the receptor-effector coupling of delta9-THC, WIN 55,212-2 and anandamide in supraspinal-induced antinociception in mice.

Impact of a mandatory syphilis delivery test on reported cases of congenital syphilis in Upstate New York.

A regulation mandating a syphilis serology test at delivery was implemented on December 6, 1989. To assess any impact on congenital syphilis reporting, we compared all cases reported in Upstate New York for the year directly preceding the regulation (n = 69) to those born in the three-year period following its implementation (n = 239). After implementation, the percentage of cases not tested at delivery decreased from 10 percent to 1 percent and reports of syphilitic stillbirths tripled. Asymptomatic infection of both mothers and babies increased significantly and at-birth detection of cases in women with negative prenatal serologies increased by 43 percent. Delivery screening failed to identify seven cases due to incubating maternal infection.

Screening criteria for Chlamydia trachomatis in family planning clinics: accounting for prevalence and clients' characteristics.

The reliability of eight self-reported risk factors as criteria for screening women for Chlamydia trachomatis was evaluated in four family planning clinics in New York State that serve diverse populations. In all, 8,920 women were screened in these clinics; the rates of infection ranged from 2% to 7%. Results of multivariate analyses showed that age was the most important predictor of chlamydial infection in the three clinics where prevalence was 4% or higher; women aged 20-24 were 3-4 times as likely as older women to be infected, and those aged 13-19 were 4-6 times as likely. In these three clinics, screening all women aged 26 or younger (62-80% of the clinic population) would identify about 90% of infected women; in the clinic with the lowest prevalence rate, age was not a reliable criterion. The prevalence of self-reported risk factors varied by clinic, and these factors generally were not reliable indicators of infection. Using the presence of at least one self-reported risk factor as a screening criterion, 80-87% of clinic clients would be screened, and about 90% of infected women would be identified. The presence of clinical signs of chlamydial infection does not increase the reliability of age as a screening criterion.

A comparative evaluation of bedside capillary blood glucose monitoring devices designed for hospital use.

Capillary blood glucose monitoring devices (CBGMs) that incorporate "wipeless" technology recently have been designed and marketed for hospital use. Our objective was to evaluate three such devices for accuracy and precision, comparing them to a popular device that utilizes older technology and to a reference standard. Blood glucose level was simultaneously determined on the CBGMs and a reference standard. Results were analyzed for precision by performing repeated measurements of a single sample and for accuracy across the entire range of determinations. Clinically relevant subsets of the entire range also were determined and arbitrarily defined as low (< 60 mg/dL), normal (60 to 140 mg/dL), high (141 to 300 mg/dL), and very high (> 300 mg/dL). We found that accuracy and precision of these devices varied considerably. Lack of accuracy was particularly evident upon analysis of the clinically relevant subset ranges of blood glucose levels. Consequently, routine evaluation of CBGMs should include analysis of clinically relevant subset ranges of blood glucose levels. The marked differences in accuracy and precision between CBGMs that are currently.

Congenital syphilis surveillance in upstate New York, 1989-1992: implications for prevention and clinical management.

Descriptive characteristics and clinical information from 322 cases of congenital syphilis were reviewed. The births (318 mothers) included 31 (10%) stillborn and 59 (18%) with clinical evidence of congenital syphilis. Only 60 (19%) had a complete laboratory workup, including radiographs of long bones and spinal fluid analysis. For a subset of 244 women with available information, 218 (89%) had > or = 1 risk factors for syphilis; however, residence in an area with high morbidity from syphilis was the only identified risk factor for 83 (34%). Eighty women (25%) were treated for syphilis during pregnancy; only 24 were treated appropriately for their stage of syphilis > 30 days before delivery. Five of these pregnancies resulted in infants with clinical signs of syphilis. These findings emphasize the need for expanded prenatal screening of high-risk women, the necessity of screening at delivery, and the need for complete evaluation of infants at risk for congenital syphilis. Further, the data suggest that in some cases therapy in the last trimester of pregnancy may be insufficient to adequately treat the fetus.

Niacin. I: Dissolution profiles of sustained-release niacin products by automated and manual procedures.

Automated and manual procedures were developed to obtain dissolution profiles of sustained-release niacin formulations. The procedures are based on the United States Pharmacopeia XXII apparatus 1 (basket) at 100 rpm with 900 mL 0.1N HCl as the dissolution medium. Filtered aliquots are read at 260 nm. No interference was found from excipients. The procedures are straightforward and will discriminate between sustained-release and regular niacin formulations.

Risk profile for Chlamydia infection in women from public health clinics in New York State.

The prevalence of chlamydial infection and associated risk factors were studied in 1531 women from ten clinics in New York State excluding New York City. Overall Chlamydia infection rates were 13.6%; 17.6% in eight high risk family planning and STD clinics, and 5.7% in two low risk college and private clinics. Risk factors for Chlamydia infection included: age < 20 years (odds ratio 1.6), use of oral contraceptives (odds ratio 2.0), a history of having more than one sexual partner (odds ratio 1.7) and, in one clinic where data was available, inflammation on Papanicolaou smears (odds ratio 2.1). These data helped secure funding for Chlamydia preventive services and permitted development of a risk profile (score card) of Chlamydia for each age group. Use of such a score card can be most helpful in assigning which patients could benefit most from Chlamydia cultures, especially in those areas where testing is unavailable or too costly to screen all patients.

Intracarotid cisplatin chemotherapy for recurrent gliomas.

Forty patients with recurrent gliomas were treated with monthly intra-arterial infusions of cisplatin. Of the 35 evaluable patients, 12 (34%) responded with computerized tomography (CT) evidence of a decrease in tumor size; in 14 (40%) the tumor stabilized on CT scans, and in nine (26%) the disease progressed. The median survival period was 35.0 weeks for the responders and 27.5 weeks for all 35 patients. The primary toxicities were renal (reversible alterations in creatinine clearance), otological (severe hearing loss in one patient), and likely neurotoxicity in one patient who had received bilateral infusions following contralateral tumor progression. The authors are now using this form of regional chemotherapy sequentially before and following radiotherapy in newly diagnosed cases.