RESUMEN

Carmi syndrome is a rare and severe disease defined by pyloric atresia and junctional epidermolysis bullosa. There are no clear recommendations when to consider a curative therapy, including surgical repair of pyloric atresia and when to transition to palliative care. We report the case of a female preterm infant suffering from Carmi syndrome. After definitive diagnosis and appropriate ethical counselling, we decided for surgical repair of the pyloric atresia. Nonetheless, there was no clinical improvement and our patient died after 35 days. Reviewing the literature, we found immunofluorescence microscopy to be most decisive examination to determine the prognosis of this severe disease.

RESUMEN

The number of patients suffering from immunodeficiency is increasing; however, the vaccination rate of these patients is below average. Administration of inactivated vaccines is harmless but does not reliably trigger a persistent immune response. Live vaccines provide a reliable protection but can cause severe disease in immunocompromised patients. Live vaccines can be administered under defined levels of immunosuppression, e.g. against measles, mumps, rubella and varicella (MMRV). In addition, the immunization of the domestic environment plays an important role in preventing infectious diseases.

Asunto(s)

RESUMEN

BACKGROUND: Respiratory infections are the main causes for hospitalization in children and a common reason for the initiation of antibiotic treatment. Rapid antigen detection tests and point-of-care mPCR-based assays provide a fast detection of viral pathogens. Nonetheless, the prescription rate of antibiotics for respiratory infections is exceedingly high. In particular, human metapneumovirus (hMPV) infections frequently cause antibiotic treatment. METHODS: Children hospitalized in our clinic with an acute respiratory infection between January 2008 and January 2013 were included in the present study. Data of 3799 children were analyzed retrospectively for clinical symptoms, laboratory findings, and antibiotic and inhalation treatment. We performed an in-house m-RT-PCR-ELISA method for pathogen detection. RESULTS: Pathogen detection was possible in 2464 patients. In 6.3%, hMPV and, in 24.0%, RSV were detected. Patients positively tested for hMPV received inhalation therapy in 62.9%; patients positive for RSV in 73.8%. Patients positive for hMPV were treated with antibiotics in 62.3%. Patients with RSV infection received antibiotic treatment in 44.4%; all others in 43.5%. Notably, a positive result in RSV-RADT was associated with reduced number of antibiotic treatment. CONCLUSION: hMPV infections inherit a two times higher probability of antibiotic treatment. There was no significant difference in laboratory findings or body temperature between hMPV infection and infections caused by other pathogens. Clinical symptoms seem not to differ from those in RSV illness. Nonetheless, RSV infections triggered significantly lower antibiotic prescription rates. A considerate application of a POC-mPCR for patients with RSV-like symptoms and age of 1 year and older with a negative RSV-RADT might lead to higher detection rates of hMPV and a reduction in prescription of antibiotics.

Asunto(s)

RESUMEN

BACKGROUND: The novel protein PTPIP51 (SwissProt accession code Q96SD6) is known to interact with two non-transmembrane protein-tyrosine phosphatases, PTP1B and TCPTP in vitro. Overexpression of the full-length protein induces apoptosis in HeLa and HEK293T cells (Lv et al. 2006). PTPIP51 shows a tissue-specific expression pattern and is associated with cellular differentiation and apoptosis in some mammalian tissues, especially in human follicular and interfollicular epidermis. PTPIP51 protein is expressed in all suprabasal layers of normal epidermis, whereas the basal layer contains PTPIP51 mRNA only but lacks the protein. OBJECTIVES: The expression of PTPIP51 was investigated in keratinocyte carcinomas, that is human basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs) as well as Bowen's disease (BD) and keratoacanthomas (KAs) on a transcriptional (mRNA) and translational (immunohistochemical) level. METHODS: Formalin-fixed, paraffin-embedded sections of BCCs, SCCs, KAs and BD, respectively, were analysed by RT-PCR, as well as immunohistochemistry and subsequent fluorescence microscopy. PTPIP51-positive cells of the tumour and the surrounding stroma were identified on the basis of specific morphological features by means of H & E staining. To obtain further information about a putative function of PTPIP51, a possible association of PTPIP51 with apoptotic cells, as well as an assumed negative correlation with proliferating cells was investigated by means of an in-situ TUNEL assay and Ki67/MIB-1 antigen staining, respectively. Co-immunostainings with PTPIP51 were performed for the following antigens: TCPTP, PTP1B and beta-catenin. RESULTS: PTPIP51-expression was detected in BCCs and SCCs of the skin, as well as in KAs and BD. Both types of keratinocyte carcinoma revealed a specific localization pattern of PTPIP51 in malignant keratinocytes. Whereas PTPIP51-positive cells of BCC were found to form two cluster types with a different subcellular localization of the protein, i.e. cytoplasmic and nuclear or predominantly membranous, investigation of SCC revealed a meshwork-like appearance of PTPIP51-positive malignant keratinocytes, created by a mainly membranous localization. BD and KA resembled the findings of PTPIP51-expression in SCC. Furthermore, we observed a partial co-localization of PTP1B and PTPIP51 in BCC. SCC and BCC showed a co-expression and partial co-localization of PTPIP51 with beta-catenin. Some PTPIP51-positive cells were found to undergo apoptosis. PTPIP51 was also expressed in cells comprising the surrounding stromal microenvironment. This was particularly noticed for endothelial cells lining peritumoural vessels as well as for infiltrating cells of both, the innate and the adaptive immune system. CONCLUSIONS: The results showed a distinct mainly membranous expression pattern of PTPIP51 in BCCs and SCCs. Since PTPIP51 was also detected in the peritumoural tissue, the protein may play a crucial role in ke

Asunto(s)

RESUMEN

High concentrations of protein tyrosine phosphatase (PTP) were found with secretory vesicles of glucagon-producing INR1G9 cells by electron microscopic immunocytochemistry, using a polyclonal antiserum specific for the PTP1B/T-cell (TC)PTP subfamily of PTP. Since TCPTP protein and mRNA were below the detection limit in the cells but significant amounts of PTP1B and mRNA were recognised by a specific monoclonal antibody and a mRNA probe we conclude, that the PTP associated with the vesicles is PTP1B. Only reverse transcriptase (RT)-PCR with primers specific for PTP1B yielded a product of the expected nucleotide sequence. Thus, we conclude that the PTP associated with the vesicles is PTP1B. The presence of vanadate for 48 h attenuated PTP1B expression and caused reduction of steady-state levels of the phosphatase. These conditions also led to a continuing increase in the steady-state rate of glucagon release by the cells. This rate and tyrosine phosphatase levels showed an inverse relationship, suggesting a suppressive role of PTP1B on the regulated secretion of glucagon by INR1G9 cells.

Asunto(s)

RESUMEN

The objective of this study was to determine the relationship between udder and leg hygiene scores of lactating dairy cattle and measures of subclinical mastitis. Study animals (n = 1250) consisted of lactating dairy cows from eight commercial dairy farms. Herds were enrolled during December 2000 and January 2001 and were visited bimonthly for a total of five visits per herd. Udder and leg hygiene scores were recorded by one person using a four-point scale ranging from one (very clean) to four (very dirty). Udder and leg hygiene scores were compared to bacteriological cultures of milk samples and monthly individual SCC values. Mean hygiene scores were 2.09 and 2.33 for udders and legs, respectively. Udder hygiene scores (UHS) were significantly associated with leg hygiene scores and varied among farms. Linear somatic cell scores increased as udder hygiene score increased. Significant differences in somatic cell scores were observed for all contrasts of udder hygiene score, except between scores of 1 and 2 and of 3 and 4. Linear somatic cell scores were associated with leg hygiene scores, but the only significant contrast was between leg hygiene scores of 2 and 4. There was a significant association between the prevalence of intramammary contagious pathogens and udder hygiene score. The prevalence of intramammary environmental pathogens was significantly associated with udder hygiene score and was 7.7, 10.0, 10.6, and 13.5% for UHS of 1, 2, 3, and 4, respectively. The prevalence of environmental pathogens was not associated with LHS. Cows with udder hygiene scores of 3 and 4 were 1.5 times more likely to have major pathogens isolated from milk samples compared with cows with hygiene scores of 1 and 2.

Asunto(s)

RESUMEN

The objective of this study was to determine the effect of tail docking on somatic cell count (SCC), intramammary infection (IMI), and udder and leg cleanliness in commercial dairy herds. Lactating dairy cows (n = 1250) from eight Wisconsin farms were blocked by farm and randomly allocated to tail docked (D) or control (C) groups. Milk samples, somatic cell counts, and hygiene scores were collected for 8 to 9 mo. The prevalence of IMI was determined for each of the five occasions when milk samples were obtained. Udder and leg cleanliness were assessed during milk sample collection. Docked and control animals were compared by logSCC, prevalence of IMI, and leg and udder cleanliness score. Variables were analyzed according to all treatment, period, and farm interactions. At the end of the study period 76 (12.2%) and 81 (13%) of cows were culled in the D and C groups, respectively. There were no significant differences in the initial data for parity, daily milk yield, logSCC, or DIM between treatment groups. Effects significant to farms were identified for all variables over all periods. Period was significant for all variables except for the prevalence of environmental pathogens, but no period x treatment interactions were detected. There was no significant difference between treatment groups for somatic cell count. The prevalence of contagious, environmental, or minor pathogens did not differ significantly between treatment groups. This study did not identify any differences in udder or leg hygiene or milk quality that could be attributed to tail docking.

Asunto(s)

RESUMEN

The primary objective of this study was to determine the behavioral and physiological effects of tail banding and atrophy using rubber rings 2 to 4 mo before first parturition in dairy heifers either with or without the use of epidural anesthesia. The secondary objective was to determine behavioral responses to tail banding using rubber rings in calves 7 to 42 d of age. Preparturient heifers (n = 24) were randomly assigned to one of four treatment groups: 1) tails were cleaned and handled; 2) tails were cleaned, handled, and an elastrator band was applied to the tail; 3) an epidural was administered 15 min before cleaning and handling; and 4) an epidural was administered 15 min before application of an elastrator band. Behavioral observations and physiological responses were collected for 6 wk. Additionally, behavioral responses to tail banding were recorded for 10 d on Holstein heifer calves that were 1 to 6 wk of age (n = 40). No significant differences in behavior were observed among treatment groups of preparturient heifers at any time during the 6-wk observation period. Preweaned calves that were 21 to 42 d of age demonstrated significantly more restlessness after application of tail bands compared to younger calves or control calves of the same age. Plasma cortisol values of preparturient heifers remained within limits previously described for nonstressed animals and no significant differences were detected among groups. Hematological values remained within the reference values for cattle, and there were no significant differences between groups except for relatively more eosinophils in the heifers that received epidurals. No significant differences in heart rate or body temperature were detected among groups.

Asunto(s)

RESUMEN

Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required to date. We now aimed at developing an effective cryopreservation method, and by testing several variables found that it was crucial to combine the most advantageous maturation stimulus with an improved freezing procedure. We generated monocyte-derived DC from leukapheresis products by using GM-CSF and IL-4 and showed that amongst several known maturation stimuli the cocktail consisting of TNF-alpha+IL-1 beta+IL-6+PGE(2) achieved the highest survival of mature DC. We then systematically explored cryopreservation conditions, and found that freezing matured DC at 1 degrees C/min in pure autologous serum+10% DMSO+5% glucose at a cell density of 10x10(6) DC/ml gave the best results. Using this approach 85-100% of the frozen DC could be recovered in a viable state after thawing (Table 1). The morphology, phenotype, survival as well as functional properties (allogeneic mixed leukocyte reaction, induction of influenza matrix or melan A peptide-specific cytotoxic T cells) of these thawed DC were equivalent to freshly prepared ones. The addition of CD40L or TRANCE/RANKL further improved DC survival. Importantly, we demonstrate that DC can effectively be loaded with antigens (such as Tetanus Toxoid, influenza matrix and melan A peptides) before cryopreservation so that it is now possible to generate antigen-preloaded, frozen DC aliquots that after thawing can be used right away. This is an important advance as both the generation of a standardized DC vaccine under GMP conditions and the carrying out of clinical trials are greatly facilitated.

Asunto(s)

RESUMEN

Dendritic cells (DCs) are considered to be promising adjuvants for inducing immunity to cancer. We used mature, monocyte-derived DCs to elicit resistance to malignant melanoma. The DCs were pulsed with Mage-3A1 tumor peptide and a recall antigen, tetanus toxoid or tuberculin. 11 far advanced stage IV melanoma patients, who were progressive despite standard chemotherapy, received five DC vaccinations at 14-d intervals. The first three vaccinations were administered into the skin, 3 x 10(6) DCs each subcutaneously and intradermally, followed by two intravenous injections of 6 x 10(6) and 12 x 10(6) DCs, respectively. Only minor (less than or equal to grade II) side effects were observed. Immunity to the recall antigen was boosted. Significant expansions of Mage-3A1-specific CD8(+) cytotoxic T lymphocyte (CTL) precursors were induced in 8/11 patients. Curiously, these immune responses often declined after the intravenous vaccinations. Regressions of individual metastases (skin, lymph node, lung, and liver) were evident in 6/11 patients. Resolution of skin metastases in two of the patients was accompanied by erythema and CD8(+) T cell infiltration, whereas nonregressing lesions lacked CD8(+) T cells as well as Mage-3 mRNA expression. This study proves the principle that DC "vaccines" can frequently expand tumor-specific CTLs and elicit regressions even in advanced cancer and, in addition, provides evidence for an active CD8(+) CTL-tumor cell interaction in situ as well as escape by lack of tumor antigen expression.

Asunto(s)

Asunto(s)

RESUMEN

In many studies the detection of p53 protein by immunohistochemistry (IHC) with one antibody is assumed to be consistent with a mutation of the gene. To determine the correlation of protein detection by immunohistochemistry and gene mutation, paraffin embedded specimens of 60 oral leukoplakias (OL) and 73 oral squamous cell carcinomas (OSCC) were analysed by IHC with two different antibodies (Do7; Pab 1801), PCR-SSCP and sequencing. In 98% of OLs and 94% of OSSCs p53 protein expression was detected with at least one antibody. Only 49% of all tissue specimen were positive with both antibodies. Molecular analysis of the same specimen showed mutations of the p53 gene in 13.3% of OLs and 9, 6% of OSCCs. p53 protein expression was not detected by IHC in 3 out of 7 OSCC with p53 mutations. According to these results detection of p53 expression by IHC is not always correlated with p53 gene mutation and failure to detect p53 protein by IHC does not prove the absence of mutation of the gene in the carcinogenesis in the oral mucosa.

Asunto(s)

RESUMEN

A supportive or causal role for human herpesvirus 6 (HHV-6) in lymphoproliferative disorders is still controversial. Different results were obtained in both tissue-based and serological investigations. We investigated 243 lymph node and salivary gland tissue biopsies for the presence of viral DNA by using a newly developed, highly sensitive nested polymerase chain reaction method. HHV-6 was detected in 39% of the non-Hodgkin's lymphomas, in 52% of Hodgkin's diseases, 64% of non-neoplastic lymph nodes, 23% of tumor metastases, and 50% of salivary gland biopsies. When correlating the patients' ages with the occurrence of HHV-6, we found a significantly higher percentage of positive samples in patients younger than 60 years of age (54%) than in older patients (35%). This age-related difference was found in all the lymphoproliferative disorders studied as well as in salivary gland biopsies. Taking patient's ages into account, we found no significant difference between the various groups of disorders concerning the percentage of HHV-6-positive samples.

Asunto(s)

RESUMEN

The unique segment of the human herpesvirus 6 (HHV-6) genome is essentially collinear to the unique long DNA segment of another betaherpesvirus, the human cytomegalovirus (HCMV). However, the HHV-6 genomic section that is analogous in position to the major immediate-early (IE) locus of HCMV does not exhibit recognizable sequence homologies. The respective HHV-6 region of 5.5 kbp is flanked on one side by 25 to 28 incomplete tandem repeats of 105 to 110 bp that contain, with one exception, a single KpnI restriction site (KpnI repeats). About 250 reiterations of the sequence motif CACATA are located on the other end. We identified two open reading frames of 375 and 2,595 nucleotides, respectively, on one strand. Strand-specific Northern blot analyses with RNA harvested from HHV-6 (strain U1102)-infected HSB-2 cells or cord blood lymphocytes revealed two transcripts of about 3.5 and 4.7 kb in the corresponding orientation. Sequence analyses of the respective cDNA clones and primer extension experiments were used to map the mRNAs. The two transcripts are coterminal and multiply spliced and code for the same putative 104.6-kDa protein, but they are initiated from different promoters. Characterization of smaller cDNA clones and Northern blotting with other strand-specific probes showed that singly spliced mRNAs of 1.0 and 1.5 kb are transcribed from the opposite strand; they could code for a 17.2-kDa polypeptide. Blocking experiments with cycloheximide led to the conclusion that only the 3.5-kb mRNA is synthesized in the absence of protein biosynthesis upon infection with cell-free virus. This identifies a single IE gene of HHV-6 at the genomic position corresponding to the major IE region of HCMV, although the coding content and transcriptional regulation are quite different for these two herpesvirus IE regions.

Asunto(s)

Asunto(s)

RESUMEN

The structural variability of the external glycoproteins of primate immunodeficiency viruses, has, so far, been investigated exclusively by sequence comparison of the respective proviral genomes. We have examined the structural relationship amount the external glycoproteins from three specific human immunodeficiency viruses (HIF-1, HIV-2), three specific simian immunodeficiency viruses from macaques (SIVmac) and three specific SIV from African green monkeys (SIVagm) by peptide mapping. Differences among glycoproteins were most pronounced between HIV-1 and SIVmac, as well as HIV-2. Two specific glycoproteins from independent SIVagm isolates were closely related to HIV-1, whereas the glycoprotein from a third SIVagm isolate was more similar to those of SIVmac and HIV-2. Our analysis reflects the classification of primate immunodeficiency viruses into three groups, the HIV-2 and SIVmac viruses, the green monkey isolates and HIV-1.

Asunto(s)

RESUMEN

The value of the oral dipyridamole-thallium stress test in identifying patients at high risk of myocardial infarction after vascular procedures has not been documented. We studied prospectively 46 patients who underwent an oral dipyridamole-thallium stress test before undergoing vascular operations. Twenty patients (43%) had a positive test result, defined by a thallium defect with reperfusion, while 26 patients had a negative test result. Myocardial infarctions were documented postoperatively in 5 (25%) of 20 of the group with positive results and 1 (4%) of 26 of the group with negative results. Three of the six myocardial infarctions were clinical; all three were in the group with positive results. No correlation was identified between dipyridamole-thallium stress test results and clinical cardiac history. A positive dipyridamole-thallium stress test result is a more sensitive predictor of postoperative myocardial infarction than ejection fraction or history of coronary artery disease. The oral dipyridamole-thallium stress test is as useful as the intravenous test in this setting.

Asunto(s)

RESUMEN

Lupus pernio has been associated with sarcoidosis of the upper respiratory tract. The semantic differences in the use of the designation "lupus pernio" are such that American dermatologists may not consider patients with nasal rim papules of sarcoidosis to have lupus pernio. During a 12-month period, we referred each new patient with sarcoidosis and nasal rim lesions for direct and indirect laryngoscopy. Three of our four patients had sarcoidosis of the upper respiratory tract. We recommend a large prospective study to assess the incidence of this disorder in patients with nasal rim lesions of sarcoidosis. Until such a study is undertaken, however, we believe that consideration should be given to otolaryngologic examination of these patients, regardless of their symptoms.

Asunto(s)

RESUMEN

The neutrophilic vascular dermatoses are a divergent group of disorders with distinct cutaneous manifestations. Diagnosis of these diseases requires clinical acumen because of the lack of pathognomonic histopathologic features. It is important for physicians to recognize these entities because of the large number of possible associated underlying diseases.

Asunto(s)

RESUMEN

Purpura is a cutaneous manifestation of a wide variety of diseases. These include such diverse entities as platelet defects, vasculitides, and disorders of connective tissue. Uncovering the underlying disorder in a patient with purpura is a stimulating challenge to the clinician's diagnostic abilities.

Asunto(s)