Granulocytic ehrlichiosis in two dogs in Switzerland.

This case report describes two dogs with granulocytic ehrlichiosis. Dog 1 was a male Labrador retriever with clinical signs of lymphosarcoma. Dog 2 was a female Airedale terrier, whose clinical signs included apathy, pyrexia, diarrhea, and abdominal pain. Examination of blood smears revealed Ehrlichia organisms in the neutrophils of both dogs. There was thrombocytopenia in both dogs, and dog 2 also had leukopenia. In both dogs, bands of identical length were amplified from DNA of leukocytes via nested PCR. The bands had identical nucleotide sequences, which differed from the gene sequences of Ehrlichia equi and E. phagocytophila in three and two positions, respectively. Interestingly, the nucleotide sequence of the 16S rRNA was 100% homologous to that of a human granulocytic ehrlichia.     

A histological and histochemical study of changes of fiber types in experimental myotonia

Summary In both fast and slow muscles of rats treated with 20–25 diazacholesterol there were qualitative alterations, such as changes of fiber outlines, numerous moth-eaten fibers and rare ring fibers. In addition there were generally larger groups of Type I and intermediate fibers than in normal controls (type-grouping tendency) in the preparations for oxidative enzymes in the extensor digitorum longus (EDL) of myotonic animals. Quantitative evaluations of EDL and soleus of myotonic rats revealed moderate hypotrophy of Type I and Type II fibers with an increase in the number of Type I and of Type III fibers in the EDL and a significant decrease of the nondominant fibers in the soleus muscle. The data are discussed in the light of a neurally mediated and/or direct action of the drug on the muscle fiber.

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Zusammenfassung Sowohl in phasischen wie auch in tonischen Muskeln von Ratten, welche mit 20–25 Diazacholesterol behandelt wurden, wurden diskrete Strukturänderungen festgestellt: Änderungen der Umrisse der Einzelfasern, zahlreiche Fasern mit Mottenfraß, seltene Ringfasern. Histochemische Reaktionen für oxidative Enzyme im Extensor digitorum longus myotonischer Tiere zeigten außerdem häufig größere Gruppen von Typ-I-Fasern und Fasern vom Zwischentyp als bei normalen Kontrollen (Tendenz zum Type-Grouping). Quantitative Auswertung zeigte im Extensor digitorum longus und im Soleus myotonischer Ratten eine mäßige Hypotrophie der Typ-I-und Typ-II-Fasern mit einer Zunahme der Anzahl der Fasern vom Typ I und vom Typ III im Extensor digitorum longus und einer signifikanten Abnahme des nicht dominanten Fasertypus im Soleus. Die Befunde werden besonders im Hinblick auf eine neurogen vermittelte oder unmittelbare Wirkung der Substanz auf die Muskelfaser diskutiert.

     

Analysis of microsatellite instability in chronic lymphoproliferative disorders

Microsatellite instability (MSI) represents one specific pattern of genomic instability and is one of the genetic lesions most frequently detected in human neoplasia. Although MSI has been found to be associated with a wide variety of solid cancers, its involvement in lymphoid malignancies is virtually unexplored. In this study, we have investigated the presence of MSI in chronic lymphoproliferative disorders by comparing the pattern of nine microsatellite repeats (two tetranucleotides, two trinucleotides, and five dinucleotides) on autologous germline and tumor DNA of 23 patients, including 17 with B-cell chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CLL/SLL), four with hairy cell leukemia, one with lymphoplasmacytoid lymphoma, and one with T-cell chronic lymphocytic leukemia. All samples at diagnosis displayed a germline pattern of the microsatellites examined, thus suggesting that MSI is not involved in the pathogenesis of these lymphoproliferations. Also, no microsatellite alterations were observed in consecutive samples of B-CLL/SLL obtained from the same patient at various stages of the disease both before and after chemotherapy. Conversely, alterations in 3/9 microsatellite repeats were detected in one case of Richter's syndrome which had evolved from a pre-existent B-CLL/SLL phase. Overall, the low frequency of MSI among chronic lymphoproliferative disorders adds further weight to the common view that the mechanisms and patterns of genomic instability in lymphoid neoplasia differ markedly from those commonly observed in solid cancers.     

The effect of improving communication competency on the certifying examination of the American Board of Surgery

BACKGROUND: Since 1991 the authors have offered a course that identifies content deficits, but only provides instruction directed at improving verbal and nonverbal behaviors. We report the outcome of this 10-year effort as success on the certifying examination of the American Board of Surgery between 1991 and 2001. METHODS: Sixteen 5-day courses were scheduled over 10 years. Participants included those who had not taken the oral examination or had failed at least once and invited senior faculty (n = 26). Sites were chosen to replicate the actual examination setting. RESULTS: There were 122 participants, with follow-up data available on 88. Success in the certifying examination after completing the course is 96 percent. CONCLUSIONS: Evaluation of communication deficits and training to improve them is strongly associated with success. Clearly, this course is effective at identifying communication behaviors that are interfering with success on the certifying examination of the American Board of Surgery.     

Initial experience using an endoscopic simulator to train surgical residents in flexible endoscopy in a community medical center residency program

INTRODUCTION: The importance of training surgical residents in GI endoscopy has been recognized for years. Despite advice from SAGES and the RRC, few programs have managed to incorporate effective flexible endoscopy training into their curriculum, making it difficult for their graduates to be credentialed in GI endoscopy. Prior to October 2001, our residents obtained their entire clinical experience in the endoscopy unit with staff surgical endoscopists. Attendance was inconsistent because of their many other responsibilities, and residents often used much of their clinical endoscopic exposure gaining basic familiarity with the equipment, precluding the development of therapeutic facility. Since October 2001, we have used the Simbionix endoscopic simulator to supplement resident training in GI endoscopy. With the advent of virtual-reality simulators, and studies validating their effectiveness in teaching fundamental technical skills, we report our initial success in implementing a formal GI endoscopy curriculum using a virtual reality endoscopic simulator to provide basic experience before the clinical endoscopic experience begins. METHODS: Residents are given monthly assignments of simulated cases on the GI Mentor simulator. Junior residents complete the diagnostic case modules; senior residents complete the therapeutic modules. Data were accumulated over the course of two years with a total of five PGY-I and eight senior surgical residents completing assigned cases on the simulator. Objective criteria were measured from their performance on the simulator to determine the efficiency of the examination for each case completed. RESULTS: Preliminary data collected over the course of two years indicates that residents improve the efficiency of their endoscopic examinations over time as measured by objective criteria. Junior surgery residents attained an aggregate average of 59% efficiency in their examinations whereas senior surgical residents who had previous experience with the simulator, attained an aggregate efficiency of 80%. CONCLUSIONS: A formal flexible endoscopy curriculum enhances surgical resident training and positively impacts careers in general and gastrointestinal surgery. Endoscopic simulators allow surgical residents to master the technical aspects of GI endoscopy quickly, thereby permitting them more benefit from their clinical exposure in the endoscopy unit. We anticipate that our formal curriculum in GI endoscopy training will prepare our graduates well for careers that include flexible endoscopy as a component of their clinical practices, and position them to be credentialled in GI endoscopy upon graduation.     

Endogenous p63 acts as a survival factor for tumour cells of SCCHN origin

The human p63 gene encodes a series of proteins that differ in their N- and/or C-terminal sequences and have widely differing properties in promoting or repressing p53-related functions such as growth arrest and apoptosis. In addition, p63 has important roles in the maintenance and differentiation of epithelial cell populations. Squamous cell carcinomas of the head and neck (SCCHN) express high levels of DeltaNp63 and p63beta isoforms compared to normal tissue from the same patients, suggesting a role for these isoforms in the pathogenesis of this common human malignancy. Here, we explore the function of p63 in SCCHN cells by using small interfering RNA (siRNA) to silence the expression of different isoforms in two SCCHN cell lines, FaDu and SCC-25. Silencing results in statistically significant decreased survival for tumour cells when all p63 isoforms, the N-terminal truncated or the alpha isoforms are inhibited. No effect was observed on cell proliferation or on the expression of epithelial differentiation markers. We also demonstrate that inhibition of endogenous p63 expression sensitises cells to the effects of ionizing radiation and cisplatin, common treatments for SCCHN patients. The data indicate that p63 has oncogenic properties in SCCHN and is predominantly involved in maintaining cell survival, rather than acting as a directly proliferative factor or as an inhibitor of terminal differentiation. Moreover, targeted inhibition of p63 expression in SCCHN could be a useful adjunct for conventional treatments of this disease.     

N-acylation of glucosamine modulates chondrocyte growth, proteoglycan synthesis, and gene expression

OBJECTIVE: To examine the effects of glucosamine (GlcN) and some N-acylated (GlcNAcyl) derivatives on the proliferation and proteoglycan (PG) synthesis of bovine articular chondrocyte (BAC); and to expand these results to human articular chondrocytes (HAC) and study the modulation of gene regulation by these compounds. METHODS: The compounds tested were: glucose (Glc), GlcN.HCl, N-acetyl GlcN (GlcNAc), and N-butyryl GlcN, (GlcNBu). GlcNBu was synthesized from GlcN and butyric anhydride. For the chondrocyte cultures, both anchorage-dependent (AD) and an anchorage-independent (AI) system (alginate beads) were evaluated. Following the various additions, BAC were assessed for total cell number, DNA, or total PG synthesis at different times. Utilizing similar conditions, human cDNA microarrays were performed for the HAC after harvesting total RNA. RESULTS: For AD cultures, the addition of GlcN.HCl (0.1-5.0 mM) to BAC or HAC cultures inhibited cell proliferation and total PG synthesis in a dose-dependent manner. For AI cultures, the inhibitory effects of GlcN.HCl on cell proliferation were less prominent, and PG synthesis increased slightly more for the GlcNAcyl than the GlcN additions. In the AD system, the addition of GlcNAc did not result in the inhibitory effect of GlcN.HCl, while GlcNBu addition resulted in an increase in BAC proliferation and PG synthesis that could not be explained by the Bu moiety alone. For the HAC, additions of 0.1 mM GlcNBu resulted in upregulation of a large number of genes, with only a few downregulated, while GlcN addition resulted in no upregulation and one downregulated gene, for preset stringency criteria. CONCLUSION: Addition of GlcNBu to BAC or HAC to AD cultures generally stimulated cell proliferation and PG synthesis, while addition of GlcN resulted in inhibition of these indicators. The inhibitory effects of the GlcN molecule appear to be related to the unsubstituted amino group. Additions of GlcNBu, but not GlcN, to HAC resulted in upregulation in the expression of a large number of genes.     

Erdheim-Chester disease: MR imaging, anatomic, and histopathologic correlation of orbital involvement

Erdheim-Chester disease (ECD) is a rare form of histiocytosis of unknown origin characterized by tissue infiltration by lipid-laden histiocytes. Typically, the diaphyseal and metaphyseal portions of the tubular bones are affected, leading to a characteristic radiographic pattern of bone sclerosis. Orbital involvement is not infrequent and is manifested by exophthalmos and periorbital xanthomatous lesions, with associated visual problems. This case report documents imaging and pathologic findings in a patient with ECD with extensive orbital involvement.     

Bone Marrow Microenvironment and the Progression of Multiple Myeloma

The BM microenvironment in MM, in terms of adhesive features, is well organized to entrap circulating precursors with BM-seeking properties and is able to produce cytokines that offer them the optimal conditions for local growth and final differentiation. Likewise, the malignant B cell clone is equipped with adhesion molecules which enable the cell to establish close contacts with BM stromal cells. Furthermore a number of cytokines are released including IL-1β and M-CSF activating ∥ BM stromal cells ∥ to produce other cytokines, such as IL-6, that stimulate the proliferation of plasma cells. Finally, most cytokines produced locally, including IL-1/β, TNF-K, M-CSF, IL-3 and IL-6, also have OAF properties, explaining why the expansion of the B cell clone parallels the activation and numerical increase of the osteoclast population.     

Afatinib in Non‐Small Cell Lung Cancer Harboring Uncommon EGFR Mutations Pretreated With Reversible EGFR Inhibitors

Background.

Afatinib, an irreversible ErbB family blocker, is approved for treatment of patients with previously untreated non-small cell lung cancer (NSCLC) harboring activating epidermal growth factor receptor (EGFR) mutations. Efficacy of afatinib in EGFR tyrosine kinase inhibitor-naïve (TKI-naïve) patients with uncommon EGFR mutations (other than exon 19 deletions or exon 21 point mutations) has been reported; however, efficacy in TKI-pretreated patients with uncommon EGFR mutations is unknown.

Materials and Methods.

In the afatinib compassionate use program (CUP), patients with advanced or metastatic, histologically confirmed NSCLC progressing after at least one line of chemotherapy and one line of EGFR-TKI treatment were enrolled. Demographic data, mutation type, response rates, time to treatment failure (TTF), and safety in patients harboring uncommon EGFR mutations were reported.

Results.

In 60 patients (63% female, median age 63 years [range: 30–84 years]), a total of 66 uncommon EGFR mutations including 30 T790M mutations were reported (18.4% and 11%, respectively, of known EGFR mutations within the CUP). Most patients (67%) received afatinib as third- or fourth-line treatment. Median TTF was 3.8 months (range: 0.2 to >24.6 months; p = .244) in patients with uncommon mutations compared with 5.1 months (range: 0.1 to >21.1 months) in patients with common mutations (n = 165). Pronounced activity was observed with E709X mutations (TTF >12 months). No new safety signals were detected.

Conclusion.

Afatinib is clinically active and well tolerated in many TKI-pretreated NSCLC patients harboring uncommon EGFR mutations. Compared with results reported in TKI-naïve patients, activity was also indicated in patients with T790M and exon 20 insertion mutations.

Implications for Practice:

This analysis consists of a large database of non-small cell lung cancer patients with uncommon EGFR mutations who were previously treated with reversible EGFR tyrosine kinase inhibitors. Although indirectly assessed, the results indicate that patients with uncommon EGFR mutations can derive benefit from treatment with the irreversible ErbB family blocker afatinib, even in some cases of tumors harboring resistance-mediating exon 20 mutations. In this study, adverse events were modest and consistent with previous reports on afatinib.