Giovanni Poletti

Molecular Characterization of a Non–Babesia divergens Organism Causing Zoonotic Babesiosis in Europe

In Europe, most reported human cases of babesiosis have been attributed, without strong molecular evidence, to infection with the bovine parasite Babesia divergens. We investigated the first known human cases of babesiosis in Italy and Austria, which occurred in two asplenic men. The complete 18S ribosomal RNA (18S rRNA) gene was amplified from specimens of their whole blood by polymerase chain reaction (PCR). With phylogenetic analysis, we compared the DNA sequences of the PCR products with those for other Babesia spp. The DNA sequences were identical for the organism from the two patients. In phylogenetic analysis, the organism clusters with B. odocoilei, a parasite of white-tailed deer; these two organisms form a sister group with B. divergens. This evidence indicates the patients were not infected with B. divergens but with an organism with previously unreported molecular characteristics for the 18S rRNA gene.

High‐dose therapy followed by autologous bone marrow transplantation (ABMT) in previously untreated non‐Hodgkin's lymphoma

13 previously untreated patients with poor prognosis non‐Hodgkins lymphoma (NHL) underwent high‐dose therapy followed by autologous bone marrow transplantation (ABMT). All patients experienced a great cytoreductive effect and 9 of them reached a complete remission (mean duration 32 months). The best results were observed in patients with more limited disease and in those without symptoms. 7 patients still remain in complete unmantained remission 15–46 months from the transplant. The probability of survival is 74% at 46 months. No therapy‐related deaths were recorded. In differentiating our preliminary approach, we propose high dose therapy followed by ABMT as induction phase in patients with stage II and as consolidation after first line therapy in patients with stages III‐IV. Further studies are warranted to determine which type of lymphoma may benefit more and which conditioning regimens may improve the remission rate.

Phase II study of a new alkylating agent (PTT-119) in resistant-relapsed non-Hodgkin's lymphomas

SummaryIn a phase II study we evaluated the effect and toxicity of a new alkylating agent, PTT-119, in 26 patients with non-Hodgkins lymphomas (NHL) resistant to or relapsed alter other chemotherapy. PTT was scheduled by ascalating the dose from 2.0 to 3.3 mg/kg every 3 weeks. Among 21 evaluable patients with NHL, 12 (57%) showed a good response (CR+PR) to PTT-119. Tolerance was acceptably good; no major side effects related to liver, cardiac, or renal toxicity were recorded. The most commonly recorded side effects were nausea and vomiting, alopecia, and phlebitis; diarrhea and drug-related fever were rarely seen. This report indicates a potential usefulness for PTT-119, a non-cross-resistant alkylating agent, in the treatment of NHL.

Babesia infection in Italy

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A rare case of Hemoglobin Leiden interfering with the DIFF channel of Sysmex XE-2100

SIR: The Hub Laboratory of The Greater Romagna Area is a ‘ Shared Resource Laboratory ’ , opened in 2009, that provides Laboratory Medicine service for more than one million inhabitants living in an area of about 5000 square km in North Italy. In the Laboratory more than 3000 Complete Blood Cell Counts (CBCs) are daily processed with a good chance of fi nding really rare cases. Recently we investigated a 37-year-old Italian woman who attended one of the 94 blood drawing centers served by our laboratory with a request of CBC in August 2014. All the hematological parameters including Red Blood Cells (RBC), White Blood Cells (WBC) and Platelets (PLT) were within reference limits but the hematology analyzer (Sysmex XE-2100) measured a very low fl uorescence signal in DIFF scattergram (Figure 1), failed to measure the differential WBC and gave the warning fl ag ‘ abnormal scattergram ’ . The surfactant applied in DIFF channel induces complete lysis and shrinkage of the RBC membrane and increases the permeability of WBC, thus permitting a polymethine fl uorescent dye to combine with nucleic acids of permeabilized cells. The intensity of fl uorescence detected by analyzer refl ects the nucleic acid content of the WBC [1]. A technical artifact appears implausible because the DIFF defect was confi rmed using the other fi ve XE-2100 analyzers equipping our laboratory, no other samples measured in that day showed the same DIFF channel defect and Quality Control yielded satisfactory results. Since the same fl uorescence DIFF defect was persistently present in three samples collected and analyzed in June 2012, April 2013 and August 2014, we suspected a stable cause of the phenomenon. We hypothesize that the RBC lysis could interfere with the WBC staining. A blood smear was observed and a normal WBC differential count with several irregularly contracted RBC were detected (Figure 2). Oxidative damage, particularly in patients with G6PDH defi ciency and some hemoglobinopathies including unstable hemoglobins, are characteristic causes of irregularly contracted RBC morphology [2]. Scandinavian Journal of Clinical & Laboratory Investigation, 2015; 75: 436–437

Critical value of laparotomy with splenectomy in stage III Hodgkin's disease as restaging procedure after chemotherapy.

38 patients with stage III Hodgkin’ disease underwent laparotomy with splenectomy as restaging procedure after first line chemotherapy which included MOPP, ABVD, or both. 28 patients were judged to be in clinical complete remission (CR) and 10 were resistant or had relapsed. Among patients in CR, 27 (96%) were confirmed to be in pathological CR; among patients resistant or relapsed, 9 (90%) were confirmed to have disease in the abdomen or retroperitoneum. The therapy for patients in clinical remission before laparotomy consisted of TNI or sTNI in 19 patients, mediastinal radiation in 6 patients and no further therapy in the remaining 3 patients. No significant differences were seen in survival and relapse‐free survival between those patients treated by extensive and those treated by local radiotherapy or no further therapy. Instead, among those patients who received extensive radiotherapy 3 developed acute non‐lymphoid leukemia (ANLL). The therapy for this group of patients consisted of further chemotherapy in 7 who had concomitant liver involvement and TNI in the remaining 3 who had the disease confined to the spleen and/or lymph nodes. Among these patients, only 3 obtained CR; 2 with radiation and 1 who was resistant to MOPP, with ABVD. This study leads us to re‐consider the role of laparotomy in stage III HD which should be used as non‐routine procedure only in selected patients without poor prognostic factors who may be cured by radiotherapy alone. In patients resistant to chemotherapy, an early evaluation of disease in the abdomen may be useful for a better salvage treatment.

Factory laboratories: Blessed are those who have been persecuted for the sake of righteousness, for theirs is the kingdom of heaven

We read with great interest the Editorial by Plebani and Lippi who discussed with the usual competence and crispness the risk that the increasing automation in clinical laboratories will outpace the laboratory professional involvement in assuring optimal tests use and interpretation [1]. According to them the “factory scenario” will induce two deleterious consequences: a progressive autonomy from the clinical context and the perception of the laboratory services as a commodity. The value of the Authors is universally recognized (their cumulated h-index is 72!) but this time we cannot agree with them. As laboratorians working in the Laboratorio Unico of Area Vasta Romagna (AVR) that includes one of the largest Corelab in Italy, a Microbiology Unit and a Genetics Unit providing laboratory testing for the citizens and the patients living in a large Area in the central Italy we wish to make a few remarks. InMarch 2009, the laboratory workload of four middle size Hospital laboratories (Cesena, Forlì, Ravenna and Rimini) and three small size Hospital Laboratories (Faenza, Lugo and Riccione) have been consolidated according to a Hub and Spokemodel with a Hub processing about 15 million tests/year and 7 spokes processing between 500,000 and 1 million tests/year. After more than 1 year of activity there are several evidences that the so called “factory laboratory” was the right and difficult answer to a (very) difficult question.