Direct printing of nanostructures by electrostatic autofocussing of ink nanodroplets.

Nanotechnology, with its broad impact on societally relevant applications, relies heavily on the availability of accessible nanofabrication methods. Even though a host of such techniques exists, the flexible, inexpensive, on-demand and scalable fabrication of functional nanostructures remains largely elusive. Here we present a method involving nanoscale electrohydrodynamic ink-jet printing that may significantly contribute in this direction. A combination of nanoscopic placement precision, soft-landing fluid dynamics, rapid solvent vapourization, and subsequent self-assembly of the ink colloidal content leads to the formation of scaffolds with base diameters equal to that of a single ejected nanodroplet. The virtually material-independent growth of nanostructures into the third dimension is then governed by an autofocussing phenomenon caused by local electrostatic field enhancement, resulting in large aspect ratio. We demonstrate the capabilities of our electrohydrodynamic printing technique with several examples, including the fabrication of plasmonic nanoantennas with features sizes down to 50 nm.

[Sequential treatment with insulin glargine and metformin, and exenatide in a patient with newly diagnosed type-2 diabetes]. / Initiale Sequenztherapie mit Insulin Glargin, Metformin und Exenatid bei Erstmanifestion eines Typ-2-Diabetes.

UNLABELLED: HISTORY AND ADMISSIONS FINDINGS: A 54-year-old man was admitted to hospital for treatment of a newly diagnosed type 2 diabetes. He had polydipsia and polyuria and had lost 11 kg in weight over four weeks. Further diagnoses were visceral obesity and arterial hypertension. INVESTIGATIONS: Laboratory tests revealed highly elevated blood glucose parameters (HbA1c 14,9 %, fasting glucose 280 mg/dl, maximal postprandial glucose 430 mg/dl and 320 mg/dl before the meal) and triglyceride (2219 mg/dl). TREATMENT AND COURSE: The patient was initially treated with a combination of insulin glargine and metformin. After a few days exenatide was added, as the patient wanted to be able to go without insulin. He was given a structured diabetes education and motivated to have five to six hours' of physical activity and a low glycemic diet. After a few days normal glucose levels had been achieved. After one month the insulin treatment was discontinued. The continuing treatment with metformin, 1000 mg twice daily, and exenatide, 5 mg twice daily, prolonged remission of beta-cell dysfunction and maintained normal blood glucose levels for seven months. After reduction of the metformin dosage (500 mg twice daily) and discontinuance of exenatide as well as a reduction of his physical activity (because of joint pain) for six months, the glucose control worsened. When exenatide was administered again, good control of postprandial blood glucose, but not of fasting glucose was achieved. CONCLUSIONS: Initial combination treatment with insulin glargine, metformin and exenatide may be beneficial in a subgroup of patients with newly diagnosed type 2 diabetes, markedly elevated blood glucose and free fatty acid levels (glucolipotoxicity). This should be further investigated in randomized controlled trials.

Postprandial blood glucose latency after oatmeal is a valid screening test for diabetic gastropathy in type 1 diabetes, but not in type 2 diabetes.

Disordered gastric motility occurs frequently in diabetes mellitus. Gastric emptying time is abnormal in about 50% of diabetic patients and delayed emptying time is known as an important cause for brittle diabetes in type 1 diabetes. We compared the rise in blood glucose after a standardized meal (oatmeal test) as a noninvasive screening test for diabetic gastropathy with the noninvasive measurement of gastric emptying time with ultrasound in type 1 and type 2 diabetic patients. The test result was considered pathological if the rise of blood glucose after an initial steady state did not reach 20 mg/dl in the first 20 min after the meal (prolonged blood glucose latency). We found a sensitivity of 90% (58.7-99.8) and a specificity of 100% (71.5-100) for the oatmeal test in type 1 diabetes in the gastropathy screening. In type 2 diabetes we found a sensitivity of 13% (1.5-38.3) and a specificity of 78% (60-90.7) (95% CI). In conclusion, the oatmeal test seemed to be a good, noninvasive screening test in diabetic gastropathy in type 1 diabetes, but has no diagnostic value in type 2 diabetes. The causes for such a difference may be due to a different postprandial blood glucose regulation in type 2 diabetes compared to the beta-cell-depleted type 1 diabetes.

[Concept of stress ulcer prevention. Is re-thinking necessary?]. / Konzept der Stressulkusprophylaxe. Ist ein Umdenken erforderlich?

BACKGROUND: The efficiency of stress ulcer prophylaxis in the prevention of gastrointestinal bleeding in critically ill patients has led to its widespread use. The lower incidence of stress ulcer bleeding, the side-effects and the cost of the prophylaxis have made it necessary targeting this preventive therapy to those patients most likely to benefit. Metaanalysis of studies on patients who received no stress ulcer prophylaxis showed few critically ill patients with important gastrointestinal bleeding. INDICATIONS: Patients who benefit most from receiving stress ulcer prophylaxis are critically ill patients with coagulopathy, or those requiring mechanical ventilation for more than two days. In patients with headinjuries, widespread burns or severe hypotension, the effects of stress ulcer prophylaxis have not been fully researched, but we would recommend administering stress ulcer prophylaxis in these cases. TREATMENT: Following a recent metaanalysis, stress ulcer prophylaxis is performed either with H2-blockers (ranitidine, famotidine) or sucralfate.

Expression of hypoxia-inducible genes in tumor cells.

Tumor tissue oxygenation impacts on proliferation of cancer cells and their sensitivity towards radio- and chemotherapy. Under low oxygen, mammalian cells show an adaptive response that leads to the induction of a number of genes with well-defined roles in oxygen supply and energy maintenance, e.g. genes encoding enzymes of the glycolytic pathway. The hypoxia-inducible factor 1 (HIF-1), a transcription factor consisting of the two proteins HIF-1alpha and HIF-1beta, plays a major role in the pleiotropic response observed under low oxygen. We have determined, by Northern analysis, the mRNA levels of HIF-1alpha and of two glycolytic enzymes known to be transcriptionally activated by HIF-1, namely phosphoglycerate kinase 1 (PGK 1) and pyruvate kinase M2 (PKM2), in different hepatoma cell lines and in mouse and human tissues. Hypoxic treatment of various mouse and human hepatoma cell lines led to the expected increase in the amount of PGK1 and PKM2 mRNA, while HIF-1alpha mRNA levels were not significantly elevated. Analysis of mouse liver tumors demonstrated no tumor-specific increases in HIF-1alpha or PGK1 mRNA levels. In five of eight human colorectal cancers investigated, PGK1 and PKM2 mRNA levels were increased in comparison to the corresponding normal tissues, while HIF-1alpha mRNA levels were not significantly changed. The majority of the colorectal cancers demonstrated p53 immunoreactivity, presumably due to mutation of the gene; there was, however, no correlation between the p53 staining pattern and mRNA expression levels of glycolytic enzymes.

Functional analysis of the human cytochrome P4501A1 (CYP1A1) gene enhancer.

The environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or dioxin) induces gene transcription, a process that requires binding of the activated aryl hydrocarbon receptor (AhR) to dioxin-responsive elements (DREs) within the enhancer region of responsive genes. Most of what is known about the molecular mechanism of AhR-dependent gene activation results from studies on the murine prototype TCDD-responsive gene cytochrome P4501A1 (CYP1A1). Much less is known, however, about the regulation of human TCDD-responsive genes. We have therefore conducted a detailed analysis of the enhancer region of the human CYP1A1 gene. From the ten DRE core motifs investigated within a stretch of 1400 bp in two human tumor cell lines using a ligation-mediated PCR technique, five motifs displayed a TCDD-inducible in vivo footprint. Four of these sites were functional enhancer sequences as demonstrated by a transient expression assay. Based on these data, a distinct functional consensus sequence for DRE motifs within the human CYP1A1 gene is suggested. After introduction of the four functional sites into various mouse hepatoma cell lines, only three exhibited a functional response, suggesting some species differences in CYP1A1 gene regulation. In addition to the footprints at DRE sites, we also detected protein-DNA interactions at three G-rich domains located within the enhancer region of the human CYP1A1 gene. Our data show that, besides some similarities in the regulation of the human and mouse CYP1A1 genes, there also exist some distinct differences, including number, location, and functional consensus sequences of DRE motifs, as well as quantity and location of footprinted G-rich domains.

Wild-type function of the p53 tumor suppressor protein is not required for apoptosis of mouse hepatoma cells.

The role of the tumor suppressor protein p53 in apoptosis of mouse hepatoma cells was studied. Different lines were used which were either p53 wild-type or carried various types of heterozygous or homozygous p53 mutations. The presence of mutations was demonstrated to correlate with a lack in transactivating activity of p53. While UV-light effectively produced apoptosis in cells of all lines, irrespective of their p53 mutational status, gamma-irradiation induced the formation of micronuclei but failed to induce apoptosis. Both UV- and gamma-irradiation led to nuclear accumulation and increases in p53 protein in p53 wild-type cells. Similarly, no significant differences in apoptotic response between p53 wild-type and p53 mutated cells were seen with other apoptotic stimuli like CD95/APO-1/Fas or TNFalpha. These data suggest that wild-type p53 is not required for induction of apoptosis in mouse hepatoma cells which may explain the apparent lack of p53 mutations in mouse liver tumors.

[Brucellosis with fatal endotoxic shock]. / Bruzellose mit tödlichem Endotoxinschock.

BACKGROUND: Brucellosis is a zoonosis with good prognosis in cases of early diagnosis. To make the diagnosis is still a problem today. CASE REPORT: A 60-year-old butcher was admitted with undulating fever, sweats, arthralgia and weight loss. Further examination revealed hepatosplenomegaly with laboratory findings of a hepatitis and multiple focal liver lesions shown by abdominal ultrasound and CT. Histologically, these lesions corresponded to caseous granulomas. Diagnosis of brucellosis was confirmed by detection of brucella species in prolonged incubation in blood culture. After the beginning of antibiotic resistance-tested therapy with tetracycline and quinolones, an endotoxic shock occurred during the first 24 hours of treatment and the patient died after multiorgan failure with disseminated intravascular coagulation. CONCLUSION: In cases of undulating fever with liver involvement, a brucellosis should be considered. Good teamwork of the internal, pathological and microbiological departments is necessary for early and correct diagnosis. This is the first report of human brucellosis in association with lethal endotoxic shock.

Cell-specific regulation of human CYP1A1 and CYP1B1 genes.

In this report, we present a characterization of the cell-specific expression of two human cytochrome P450 genes, CYP1A1 and CYP1B1, by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The TCDD-dependent induction of CYP1A1 has been studied extensively and serves as the prototype response for a TCDD-signaling pathway initiated by the reversible binding of TCDD to an intracellular receptor [designated the aryl hydrocarbon (Ah) receptor]. CYP1A1 is induced by TCDD to high levels (45-fold increase) in the human hepatoblastoma line HepG2 as compared with the human renal adenocarcinoma line ACHN. In contrast, CYP1B1 is induced selectively in ACHN cells. Cell-specific induction of CYP1A1 and CYP1B1 mRNA correlates with comparable changes in the corresponding proteins and results, at least in part, from transcriptional activation. Characterization of the mechanism(s) for the differential regulation of CYP1A1 was carried out. Nuclear extracts obtained from either cell line following treatment with TCDD displayed equivalent binding to oligonucleotide probes for two dioxin-responsive elements located 5'-ward of the CYP1A1 promoter. This result obtained with broken cell fractions was confirmed by an intact cell DNA protection assay. Possible involvement of negative regulators is suggested by the presence of a negative regulatory element in the 5' flanking region of the CYP1A1 gene and the observed superinduction of CYP1A1 mRNA by cycloheximide in TCDD-treated HepG2 cells. Electromobility shift analysis using negative regulatory element probes, however, did not detect quantitative differences in the binding of nuclear extract proteins obtained from either HepG2 or ACHN cells treated with TCDD. These findings indicate that the ligand-dependent activation and dioxin-responsive element binding of the Ah receptor required for CYP1A1 induction in HepG2 cells also can occur in ACHN cells. We conclude that the repression of TCDD-dependent CYP1A1 induction in ACHN cells occurs at the level of transactivation in the Ah receptor signal transduction pathway.

[Long-term results of endoscopic balloon dilatation of ulcer-induced pyloric stenoses--follow-up of 25 patients]. / Langzeitergebnisse der endoskopischen Ballondilatation ulkusbedingter Magenausgangsstenosen--Follow-up von 25 Patienten.

Long-term results of the "through the scope balloon dilatation" for treatment of benign pyloric stenosis are lacking. Therefore we retrospectively analyzed 25 patients treated by balloon dilatation because of benign pyloric stenosis between 1986 and 1995. The mean age was 63 years (18-88 years), there were twelve men and 13 women. The mean follow-up period was 38 months (twelve to 120 months). 92% (23 patients) underwent successful dilatation. 20% (five patients) presented with recurrent obstruction. They were treated again and none of them has had symptoms up to now. There were no complications due to dilatation. Through the scope balloon dilatation is a successful and cost effective therapy for benign gastric outlet obstruction also in long-term follow-up.

Dysphagia secondary to left atrial dilatation.

Dysphagia, of a cardiac origin, is a generally accepted, yet rarely diagnosed symptom. A 84-year-old female patient with left atrial dilatation, presenting with dysphagia and weight loss, was treated for global heart failure. Esophagography revealed compression of the distal esophagus. Echocardiography showed a left atrial, right atrial and right ventricular enlargement. Esophageal manometry revealed hypotonic peristaltic contractions of the esophageal body and a lower than normal resting pressure of the lower esophageal sphincter. After medical treatment for myocardial insufficiency and of esophageal motility, the clinical signs of global heart failure were improved and the dysphagia disappeared. Following the exclusion of common causes of dysphagia in patients with heart failure, dysphagia due to left atrial dilatation should be considered.

Primary bilateral renal centrocytic non-Hodgkin's lymphoma as a cause of renal failure.

We present a patient whose cause of renal failure was primary and isolated bilateral renal manifestation of centrocytic non-Hodgkin's lymphoma. The treatment options for bilateral primary renal lymphoma are discussed against the background of published data concerning this topic.

Gamma-irradiation-induced micronuclei from mouse hepatoma cells accumulate high levels of the tumor suppressor protein p53.

The p53 tumor suppressor protein plays an important role in the G1 arrest of cells treated with DNA-damaging agents. Mouse hepatoma cells with wild-type or mutated p53 genotype were gamma-irradiated and the time course of p53 expression was determined by immunocytochemical staining. In p53 wild-type cells, gamma-irradiation led to a transient accumulation of the protein in the nuclei, whereas no such accumulation occurred in p53-mutated cells. Micronuclei were induced by gamma-irradiation in both wild-type and mutated cells in a dose- and time-dependent manner, but only micronuclei from p53 wild-type cells demonstrated a strongly positive staining reaction for p53 protein. This accumulation of p53 protein in micronuclei was not associated with a block in DNA synthesis as evidenced by bromodeoxyuridine labeling experiments.

[Acute pancreatitis due to the rupture of an echinococcal cyst into the bile duct system]. / Akute Pankreatitis infolge Einbruchs einer Echinococcuszyste in das Gallengangssystem.

A 47-year-old woman with stones in the gall-bladder suddenly developed severe upper abdominal pain. Cholesterol concentration was elevated, as were amylase (555 U/l) and lipase (408 U/l) concentrations, suggesting biliary pancreatitis. Endoscopic retrograde cholangiography demonstrated a cyst, about 10 cm in diameter, in the left lobe of the liver, connected to the biliary tract system. Ultrasonography and computed tomography additionally showed a smaller cyst in the right lobe. Infection with Echinococcus granulosus was proven microbiologically on bile (demonstration of hooklets and protoscolices) as well as serologically. Transpapillary cholangioscopy demonstrated daughter cysts within the echinococcal cyst. The main cyst was rinsed with 20% NaCl for 10 days via a nasocystic catheter. In addition, mebendazole (three times daily 1000 mg) was administered for 13 months. The signs if inflammation receded and the cyst shrank to a small residual volume. Surgical intervention became unnecessary.

[Detection of the IgE receptor CD23 in sarcoidosis]. / Nachweis des IgE-Rezeptors CD23 bei Sarkoidose.

In 30 sarcoidosis patients the soluble IgE receptor sCD23 was determined. Compared to healthy controls a marked increase of this molecule could be detected in serum (4.9 versus 0.9 ng/ml). In 19 patients who underwent bronchoscopy a strong expression of CD23 on alveolar macrophages could be demonstrated in immunochemistry compared to healthy controls (40.8 versus 12.6%). Determination of total IgE and anti-IgE antibodies did not allow discrimination between healthy controls and sarcoidosis patients. Our data suggest that alveolar macrophages are an important source for soluble CD23 in serum. CD23 expression might reflect functional activation of macrophages since our results do not appreciate a role for IgE-related reactions in sarcoidosis.

Discrimination between periportal and pericentral necrosis of rat liver by determination of glutamine synthetase and other enzyme activities in serum.

Periportal or pericentral necrosis of rat liver was produced by injection of allyl-alcohol or bromobenzene, respectively. Activities of predominantly periportal and perivenous enzymes were determined in serum during maximal necrosis. Aspartate aminotransferase, which is more or less homogeneously distributed in the liver acinus, exhibited similar activities in serum after periportal and pericentral injury. Serum activities of the mainly periportal enzymes alanine aminotransferase and fructose 1,6-bisphosphatase were 1.5- to 2-fold higher after periportal as compared to pericentral necrosis. Serum activity of the mainly pericentral glutamate dehydrogenase was 3-fold higher after pericentral than after periportal damage. However, due to individual variations necrosis could not be definitively localized in any case by measurement of these enzyme activities. Better discrimination between periportal and pericentral necrosis was achieved by the serum activity of the exclusively pericentral enzyme glutamine synthetase, which was 8-fold higher after pericentral as compared to periportal necrosis. Conclusive discrimination was obtained by the activity ratio fructose 1,6-bisphosphatase/glutamine synthetase in serum.

Expression of the adhesion molecule ICAM-1 on alveolar macrophages and in serum in extrinsic allergic alveolitis.

ICAM-1 expression was studied on alveolar macrophages (AM) in extrinsic allergic alveolitis (EAA) using immunocytochemistry and cytometry. Compared to control donors, EAA patients had higher percentages of ICAM-1+ AM (75% vs. 30%) and the antigen density was more than sixfold higher (673 vs. 103 channels). Levels of soluble serum ICAM-1 were found increased in EAA patients with an average of 866.2 +/- 300 ng/ml versus 394.8 +/- 110 ng/ml in controls. These data are consistent with the concept that antigen contact upregulates ICAM-1 expression on AM in EAA followed by shedding and an increase in serum sICAM-1.

Carcinogen-specific mutational pattern in the p53 gene in ultraviolet B radiation-induced squamous cell carcinomas of mouse skin.

We have examined 35 epidermal tumors induced in mice of four different strains by chronic exposure to ultraviolet B radiation for the presence of aberrations in the p53 tumor suppressor gene. Polymerase chain reaction products from p53 exons 5 to 8 were screened by single-strand conformation polymorphism analysis and sequencing. Base substitutions were found in seven tumors (20%). All mutations occurred at dipyrimidine sequences; most frequent were C-->T single base and CC-->TT tandem transitions suggesting the involvement of UV radiation in the genesis of the mutations. Three base substitutions were located at codon 148, and all dipyrimidine-derived mutations occurred at sites where the sequence is present in the nontranscribed DNA strand, indicating some site and strand specificity of the ultraviolet B-induced p53 mutations.