Sacral nerve stimulation (SNS), posterior tibial nerve stimulation (PTNS) or acupuncture for the treatment for fecal incontinence: a clinical commentary.

Sacral nerve stimulation (SNS) has become an established therapy worldwide for the treatment for fecal incontinence. A large number of papers have been published over the years, and SNS is generally considered very effective with improved continence and quality of life for most patients. However, the results are mostly expressed in the semi-quantitative terms, that is, patients' diaries translated into score points. The clinical value of SNS is questionable, especially as the patient groups are usually small and/or etiologically heterogenic and the follow-up period mostly short. The Health Technology Assessment organization in the west region of Sweden has recently evaluated the SNS with regard to evidence, efficacy and risks. Economic and ethical aspects raise serious questions on this expensive and not entirely risk-free treatment in routine medical care. Similar criticism has also been raised by other reviewers proposing a more thorough scientific assessment with well-designed randomized trials and comparison with other similar methods of treatment.

A comparison of three diagnostic techniques for malaria: a rapid diagnostic test (NOW Malaria), PCR and microscopy.

Malaria is a common, life-threatening infection in endemic tropical areas and one that presents a diagnostic challenge to laboratories in most non-endemic countries. A rapid and accurate diagnosis is a prerequisite for effective treatment, especially for the potentially fatal cases of Plasmodium falciparum infection. In the present, multi-centre study, the performances of a rapid diagnostic test (NOW) Malaria) and several, commercial, PCR-based assays (AMS61, AMS42, AMS43, AMS4 and AMS45) were compared against the results of microscopical examination of bloodsmears (the current 'gold standard'). The subjects were either non-European immigrants (N=135) or international travellers (N=171). There was good concordance between the results of all the detection methods, with kappa values of >0.8. Although the NOW Malaria rapid test was both sensitive (100%) and specific (100%) in detecting P. falciparum infections, it was less specific (93.1%) and sensitive (90.7%) in identifying the other Plasmodium species. The results from the AMS61 assay, designed to detect any malarial infection, generally parallelled those of the microscopy (kappa = 0.89), giving a specificity of 98.2% and a sensitivity of 91.0%. Although the use of species-specific molecular primers to identify pure infections with P. falciparum and P. vivax gave results that were in good agreement with those of the microscopy, the subjects who had apparently pure infections with P. ovale or P. malariae were always found PCR-negative. Compared with the standard microscopy, both the NOW Malaria test and the PCR-based assays were therefore poor at identifying mixed infections. The NOW Malaria test and the PCR-based assays clearly need to be improved, particularly for the correct identification of infections with Plasmodium spp. other than P. falciparum, including mixed infections. For now, expert microscopy must remain the mainstay of the laboratory diagnosis of malaria.

Is an ileal pouch an alternative for patients requiring surgery for Crohn's proctocolitis?

BACKGROUND: Most surgeons consider Crohn's colitis to be an absolute contraindication for a continent ileostomy, due to high complication and failure rates. This opinion may, however, be erroneous. The results may appear poor when compared with those after pouch surgery in patients with ulcerative colitis (UC), but the matter may well appear in a different light if the pouch patients are compared with Crohn's colitis patients who have had a proctocolectomy and a conventional ileostomy. METHODS: We assessed the long-term outcomes in a series of patients with Crohn's colitis who had a proctocolectomy and a continent ileostomy (59 patients) or a conventional ileostomy (57 patients). The median follow-up time was 24 years for the first group and 27 years for the second group. RESULTS: The outcomes in the two groups of patients were largely similar regarding both mortality and morbidity; the rates of recurrent disease and reoperation with loss of small bowel were also similar between groups. CONCLUSIONS: The possibility of having a continent ileostomy, thereby avoiding a conventional ileostomy-even if only for a limited number of years--may be an attractive option for young, highly motivated patients.

The ileoanal pouch procedure in the long-term perspective: a critical review.

An ileo-pouch anal anastomosis (IPAA) has become the gold standard procedure for ulcerative colitis and familial adenomatous polyposis. Clinical results on the pelvic pouch procedure have often been encouraging; when confronted with the different surgical options, the majority of patients select IPAA as the best operation. However, even if IPAA is a great innovation, it is by no means the first choice for all patients. For patients old enough to join in a responsible discussion, the pros and cons of the various operations must be carefully described; the choice of surgical procedure must meet the patient's wishes and appear soundly based to the surgeon. The young age of most patients has to be considered and a long follow-up time is required to establish whether and, if so, to what extent the operation may adversely impact the patient's continence, sex life, fertility, and quality of life. The risk of cancer transformation in the residual rectal mucosa in the muscular or columnar cuff is another important factor that may influence the eventual decision. This article critically reviews our experience and the literature.

Oviposition sequence and offspring of mated and virgin females of Cotesia flavipes (hymenoptera: braconidae) parasitizing Diatraea saccharalis larvae (lepidoptera: crambidae)

Large scale mass rearing of natural enemies has been a mean of improving biological control in the sugarcane intensive agriculture. Among them, Cotesia flavipes, a gregarious koinobiont endoparasitoid, was imported by Brasil to control caterpillars of the sugarcane borer Diatraea saccharalis. The C. flavipes larval development depends on its association with polydnavirus, which blocks the host defense reaction. To verify if the oviposition sequence (1st, 2nd or 3rd) and the female condition (mated or virgin) interfere in the number of C. flavipes descendents, 4th instar caterpillars of D. saccharalis were parasitized. Analysis of the data showed that: a) there is an inverse correlation between the parasitism efficiency and the host reaction (encapsulation); b) the number of caterpillars parasitized by virgin females that released parasitoid larvae in the period from 12 to 15 days was higher than that of caterpillars parasitized by mated females; c) a slight difference between mated and virgin females in relation to the parasitim success was observed; and d) the number of encapsulated parasitoid larvae was higher than that of eggs, suggesting that eggs have a better capacity to overcome the host reaction. In this study, the viability of C. flavipes eggs and larvae in the non-specific host D. saccharalis could be correlated with the oviposition sequence and the female condition

Diagnosis of visceral leishmaniasis: the sensitivities and specificities of traditional methods and a nested PCR assay.

In the present study, 67 patients suspected to be cases of visceral leishmaniasis (VL) were each checked for leishmanial infection by the microscopical evaluation of various biological specimens, in-vitro culture, serology and an assay based on nested PCR. Most (35) of the subjects were immunocompetent (IC) but 32 were immunodeficient (ID) as the result of HIV infection (18 cases), treatment to prevent transplanted organs being rejected (six) or haematological malignancies (eight). Forty-one (61.2%) of the subjects (19 IC subjects, 12 HIV-positive patients, four transplant patients and six patients with malignancies) were considered true cases of VL. For the IC subjects, only the production and microscopical examination of leucocytoconcentrates and cultures of Buffy coats gave sensitivities of <80%, the results of the other methods showing higher sensitivities and almost perfect agreement with the 'gold-standard' diagnoses. For the ID subjects, however, only the serological tests and the PCR gave reasonable sensitivities (of >80%). For the initial diagnosis of leishmaniasis in ID patients, IFAT and western blots may be useful, as, among the present ID patients, they gave sensitivities (of 80.9% and 88.2%, respectively) that were almost as high as that for the PCR, and specificities of 100%. In the diagnosis of VL in either IC or ID patients, the assay based on a nested PCR appeared to be particularly reliable, with sensitivities of 88.9% and 95.2%, respectively, and a specificity of 100% in both groups of patients. The testing of bone-marrow aspirates by PCR revealed very few VL cases who were not found positive when samples of their peripheral blood were checked in the same assay. For both IC and ID subjects therefore, the use of the PCR-based method to test samples of peripheral blood (which can be collected much more easily than bone-marrow aspirates and with much less pain for the subject) is recommended.

Haemolymph electrophoretic pattern of Ascia monuste orseis larvae (Lepidoptera: Pieridae) parasitized by Cotesia glomerata (Hymenoptera: Braconidae)

Cotesia glomerata is a natural enemy of the vegetable plague Ascia monuste orseis and preferably parasites 2nd, 3rd and 4th instar larvae. Parasitism effects on the haemolymph protein profile of Ascia monuste orseis larvae from the 2nd to 7th days were analyzed both qualitatively and quantitatively by SDS-PAGE and Coomassie-Blue binding methods. Quantitative analysis showed a progressive increase in the protein content of about 6.5 and 12.5 times in parasitized and non-parasitized larvae from the 2nd to 5th days, respectively. On the 6th day, a decrease in protein content was observed in both groups, although this decrease was significantly less than the control group that continued to metamorphosis. Meanwhile, parasitized larvae had one more day (7th day) in their larval period to complete parasitoid development, justified by the fact that parasitoid is koinobiont and allows host feeding. On this day, a drastic increase in protein content was detected when the parasitoids left the host. The SDS-PAGE showed proteins of high molecular weight (>120 kDa) on the 5th day of the non-parasitized larvae when they entered pre-pupa stage and on the 7th day of parasitized larvae. Proteins with MW lower than 62 kDa and higher than 27 kDa were absent on the 5th day in control larvae (pre-pupa phase), but present in parasitized larvae. This could indicate a possible relation between these proteins and the host juvenile hormone. Therefore, the presence of C. glomerata influences Ascia monuste orseis development, but its own physiological development is apparently independent of the host, which tends to die when parasitism succeeds.

Multicentre study, in patients with imported malaria, on the sensitivity and specificity of a dipstick test (ICT Malaria P.f./P.v.) compared with expert microscopy.

A prospective, multicentre study was carried out in Italy to assess the sensitivity and specificity of a rapid dipstick test (ICT Malaria P.f./P.v.) in the diagnosis of imported malaria caused by Plasmodium falciparum and other Plasmodium spp. The test is based on the detection of histidine-rich protein-2 (HRP-2) from P. falciparum and 'panmalarial' antigen in peripheral blood. The 241 subjects were international travellers or immigrants from areas where malaria is endemic. When compared with the microscopical examination of bloodsmears (used as the 'gold standard'), the dipsticks were found to be 94.4% sensitive and 94.5% specific for pure infections with P. falciparum. The performance of the tests when used on patients infected with species other than P. falciparum or more than one Plasmodium spp. showed a high degree of variability. Although the dipsticks represent a very simple, rapid, and valuable diagnostic aid, they should not be considered a complete substitute for direct microscopical diagnosis using stained bloodsmears.

Genetic diversity in the microsporidian Encephalitozoon hellem demonstrated by pulsed-field gel electrophoresis.

Encephalitozoon hellem is a microsporidian species responsible for opportunistic infections in AIDS patients. Use of a novel chitinase-based method allowed unsheared chromosomal DNA to be recovered from eleven E. hellem isolates derived from three geographic regions. All isolates were typed by 18S rDNA sequencing, which showed that they belonged to intemal transcribed spacer type 1. After ethidium bromide staining, pulsed-field gel electrophoresis (PFGE) analysis discriminated two new karyotypes comprising 7 and 8 chromosomal bands respectively, ranging in size from 205- to 272-kb pairs. Genomic size was estimated to be 2.39 Mb. Our data indicate PFGE is useful for typing E. hellem and confirms genetic diversity among E. hellem genotypes.

Reappearance of human diphyllobothriasis in a limited area of Lake Como, Italy.

BACKGROUND: Very few sporadic cases of autochthonous diphyllobothriasis have been reported in Italy during the past 20 years, probably due to the drastic decrease or disappearance of some fish species, mainly Perca fluviatilis L., known as the intermediate host of the infective larval stage of this tapeworm. However, increasing consumption of raw or undercooked fish during the last years as influenced by Oriental cuisine, played an important role in a number of new single cases of the infection or microepidemics. PATIENTS: We describe six cases which were diagnosed within a period of 4 months, which all originated from the area of Lake Como (Lombardy region). All subjects had previously eaten raw perch fillets. Four individuals complained of gastrointestinal signs and symptoms, whereas two were asymptomatic. DISCUSSION: Parasitological diagnosis was achieved in four cases by recovery of typical operculated eggs in stool specimens; in two others by morphological features of proglottids recovered from feces. Treatment with niclosamide, as a single oral dose, cured the infection in all subjects. This is the first report of this infection in Italy in the last 10 years.

Genotyping Encephalitozoon hellem isolates by analysis of the polar tube protein gene.

To develop an alternative genotyping tool, the genetic diversity of Encephalitozoon hellem was examined at the polar tube protein (PTP) locus. Nucleotide sequence analysis of the PTP gene divided 24 E. hellem isolates into four genotypes, compared to two genotypes identified by analysis of the internal transcribed spacer of the rRNA gene. The four PTP genotypes differed from each other by the copy number of the 60-bp central repeat as well as by point mutations. A simple PCR test was developed to differentiate E. hellem genotypes based on the difference in the size of PTP PCR products, which should facilitate the genotyping of E. hellem in clinical samples.

Injection treatment of hemorrhoids in patients with acquired immunodeficiency syndrome.

PURPOSE: Patients with acquired immunodeficiency syndrome are often in poor general physical condition. Diarrhea and bleeding hemorrhoids frequently contribute to the morbidity, and patients with such problems cause an increasing load on many outpatient clinics. METHODS: Twenty-two patients (17 males) with acquired immunodeficiency syndrome had injection treatment for bleeding second-degree to fourth-degree hemorrhoids according to standard outpatient clinic routines. Mean follow-up was 24 months. RESULTS: No complications were recorded. The treatment was successful in all patients, and no hemorrhoidectomy was necessary. Nineteen patients improved after their first injection, whereas 3 patients required two to six weeks repeated treatments to improve. Four subjects with the longer follow-up (4 years) showed an improvement lasting 12 to 18 months and then required one to two treatments per year to stop recurrent bleeding. CONCLUSIONS: Because of their poor general condition and poor wound healing, a conservative approach is preferable to avoid a formal hemorrhoidectomy in patients with acquired immunodeficiency syndrome. Sclerotherapy seems to be an attractive alternative.

[Pathogenic protozoans in man: differential characterization of Entamoeba histolytica/Entamoeba dispar complex, Acanthamoeba spp., Microsporidia]. / Protozoi patogeni per l'uomo: protocolli di caratterizzazione differenziale di genere e di specie nell'ambito del complesso Entamoeba histolytica/Entamoeba dispar, Acanthamoeba spp., Microsporidi.

The review summarizes the results in the main parasitological topics of our Lab: amoebic infections due to Entamoeba histolytica/Entamoeba dispar complex and to Acanthamoeba spp. respectively, and human infections caused by microsporidia. Different rapid and advanced techniques have been included in the standardized diagnostic protocols for each topic, and a critical comparison among them was made, in order to define the gold standard diagnostic method: a) E. histolytica/E. dispar: in vitro culture, zymodeme typization, biomolecular identification (PCR), immunoenzymatic assay (ELISA) for direct detection in stools of specific surface antigenic lectins; b) Acanthamoeba spp.: in vitro culture, light and ultrastructural characterization, species identification by immunofluorescence method with monoclonal antibodies, in vitro pharmacological studies; c) Microsporidia: ultrastructural (TEM), biomolecular (PCR), biochemical and immunological (SDS-PAGE, Immunoblotting) studies for species identification, use of advanced ultrastructural techniques ("freeze-etching", "deep-etching") in order to deepen the spore wall structure, to study the cytoskeletal function of actin and to define the mode of infection, in vitro pharmacological assays on some inhibitors of chitin-synthases.

Asymptomatic amoebic infection: Entamoeba histolytica or Entamoeba dispar? That is the question.

Two cases of amoebic infection were diagnosed in a heterosexual couple. The cases, a Frenchman with previous trips to various African countries and his sexual partner, a Cameroonese woman immigrant living outside the community, were both asymptomatic; the infection had been diagnosed by chance in the man at the time of his employment in a hospital kitchen. Based on what is known of the epidemiology of amoebic infection, it may be acquired and then transmitted within a couple via the indirect faecal-oral route or, in greater likelihood, by sexual practices. Both amoebic isolates were characterised by isoenzyme electrophoresis as non-pathogenic Entamoeba dispar, zymodemel. Other diagnostic tools, such as ELISA direct stool antigen detection tests and serological assays were employed, confirming the diagnosis of E. dispar infection. Given there are a number of asymptomatic cyst passers of Entamoeba histolytica, besides human carriers of saprophyte E. dispar, we stress the importance of applying, when possible, advanced protocols of diagnosis to distinguish the microscopically identical pathogenic species from the non-pathogenic one.

In vitro efficacy of nikkomycin Z against the human isolate of the microsporidian species Encephalitozoon hellem.

Since 1985 microsporidia have been recognized as a cause of emerging infections in humans, mainly in immunocompromised human immunodeficiency virus-positive subjects. As chitin is a basic component of the microsporidian infective stage, the spore, we evaluated in vitro the susceptibility of a human-derived strain of Encephalitozoon hellem to nikkomycin Z, a peptide-nucleoside antibiotic known as a competitive inhibitor of chitin synthase enzymes. Transmission electron microscopy showed that this drug, at 25 microgram/ml, reduced the number of parasitic foci by about 35% +/- standard deviation after 7 days of culture (P < 0.0001) and induced cell damage of both mature and immature spores and also other sporogonic and merogonic stages. In particular, an irregular outline of the cell shape and an abnormally condensed cytoplasm in meronts and sporonts were documented. Also, the polar tubule and the polaroplast membranes appeared disarrayed in the sporoblast stage. The spore wall showed an enlarged endospore and delaminated exospore. Mature spores had a complete cytoplasmic disorganization and a swollen and delaminated cell wall. No ultrastructural cell damage was observed in uninfected control cultures treated with the drug.