Inoculum effect of CTX-M-15, OXA-48, and KPC-2 producing Klebsiella pneumoniae on meropenem and ceftazidime-avibactam efficacy.

The inoculum effect, characterized by diminished antibacterial activity at high bacterial inocula, is studied in the context of beta-lactam and beta-lactamase inhibitor combinations against beta-lactamase-producing Enterobacterales. The inhibition of ESBL + OXA-48 and KPC enzymes, in combination with ceftazidime, demonstrates encouraging results. In this study, 20 Klebsiella pneumoniae isolates were tested with different inocula (1-5 × 105 and 1-5 × 107 cfu/ml) using broth microdilution methods. The inoculum effect was observed in meropenem against OXA-48 + CTX-M-15- and KPC-2-producing isolates but not with ceftazidime/avibactam. Notably, meropenem exhibited inoculum effect against carbapenemase-producing strains, whereas ceftazidime-avibactam remained effective. We conclude that ceftazidime-avibactam is recommended for high-inoculum infections.

Surface electrical stimulation of the auditory cortex preserves efferent medial olivocochlear neurons and reduces cochlear traits of age-related hearing loss.

The auditory cortex is the source of descending connections providing contextual feedback for auditory signal processing at almost all levels of the lemniscal auditory pathway. Such feedback is essential for cognitive processing. It is likely that corticofugal pathways are degraded with aging, becoming important players in age-related hearing loss and, by extension, in cognitive decline. We are testing the hypothesis that surface, epidural stimulation of the auditory cortex during aging may regulate the activity of corticofugal pathways, resulting in modulation of central and peripheral traits of auditory aging. Increased auditory thresholds during ongoing age-related hearing loss in the rat are attenuated after two weeks of epidural stimulation with direct current applied to the surface of the auditory cortex for two weeks in alternate days (Fernández del Campo et al., 2024). Here we report that the same cortical electrical stimulation protocol induces structural and cytochemical changes in the aging cochlea and auditory brainstem, which may underlie recovery of age-degraded auditory sensitivity. Specifically, we found that in 18 month-old rats after two weeks of cortical electrical stimulation there is, relative to age-matched non-stimulated rats: a) a larger number of choline acetyltransferase immunoreactive neuronal cell body profiles in the ventral nucleus of the trapezoid body, originating the medial olivocochlear system.; b) a reduction of age-related dystrophic changes in the stria vascularis; c) diminished immunoreactivity for the pro-inflammatory cytokine TNFα in the stria vascularis and spiral ligament. d) diminished immunoreactivity for Iba1 and changes in the morphology of Iba1 immunoreactive cells in the lateral wall, suggesting reduced activation of macrophage/microglia; d) Increased immunoreactivity levels for calretinin in spiral ganglion neurons, suggesting excitability modulation by corticofugal stimulation. Altogether, these findings support that non-invasive neuromodulation of the auditory cortex during aging preserves the cochlear efferent system and ameliorates cochlear aging traits, including stria vascularis dystrophy, dysregulated inflammation and altered excitability in primary auditory neurons.

Proposal of model for personalized early adapted cancer screening in people living with HIV: experience of "Gaetano Martino" Hospital University of Messina.

Human immunodeficiency virus (HIV) infection has historically been related to the development of specific cancers, some of which are so closely linked to the infection, such as Kaposi's Sarcoma (KS), that they have earned the name Acquired Immuno-Deficiency Syndrome (AIDS)-defining cancers (ADCs). While the development of antiretroviral therapy (ART) has decreased the incidence of AIDS-defining cancers, the resulting aging of people living with HIV (PLWH) highlighted an increased occurrence of other forms of cancer. At the "Gaetano Martino" hospital in Messina, we developed a multidisciplinary approach by creating a bridge between the Oncology Unit and the Infectious Diseases Unit to carry out screening and a more rapid diagnostic and therapeutic journey for cancers in PLWH. The goal is to improve the diagnosis of various types of cancer by involving other professionals, such as gastroenterologists and gynecologists, to ensure faster access to treatment and, therefore, a greater chance of survival. In addition, our multidisciplinary approach has also included vaccine screening, offered by the "Gaetano Martino" hospital and useful for preventing the development of specific forms of cancer in the entire population and particularly in PLWH.

A comparative study of two different methods of sample preparation for positive blood cultures for the rapid identification of bacteria using MALDI-TOF MS.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has lately been implemented as a solid technology for rapid microorganism identification in microbiology laboratories. This study compares two methods for bacterial separation from 85 positive blood culture before MALDI-TOF MS: (1) a conventional method that we used in our laboratory to prepare bacteria for susceptibility testing and (2) a new commercialized technique (Sepsityper). There were no significant differences in the identification of Gram-negative bacilli regardless of the bacterial separation method used. However, identification was greater for Gram-positive cocci when the Sepsityper method was used (84.15% vs. 100% in the identification to a genus level in staphylococci and 57.14% vs. 85.71% in the identification to a genus level of enterococci with the in-house and Sepsityper methods, respectively). Therefore, the Sepsityper method to prepare bacteria from a positive blood culture is more adequate for the further identification of Gram-positive cocci by MALDI-TOF MS.

Plasmapheresis for treatment of pulmonary alveolar proteinosis.

Whole lung lavage (WLL) is currently the standard therapy for pulmonary alveolar proteinosis (PAP). Nevertheless, some PAP patients respond poorly to WLL or require it frequently. The present paper reports a patient with autoimmune PAP with persistent disease despite three WLL treatments over 10 months. Plasmapheresis with ten 1.5-L plasma exchanges was performed, which lowered the serum granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibody level from 250 microg mL(-1) to 156 microg mL(-1) but did not improve respiratory impairment. Further WLL therapy was required and transiently effective. Serum GM-CSF autoantibody levels declined progressively, reaching a value of 56 microg mL(-1) 80 weeks after completion of plasmapheresis. However, this decrease was not accompanied by clinical improvement and the patient required additional WLL therapy. The results confirm that minor reductions in serum granulocyte-macrophage colony-stimulating factor autoantibody levels from plasmapheresis are not reflected in clinical improvement in the severity of lung disease in pulmonary alveolar proteinosis.

Secondary outputs of alpha1-antitrypsin deficiency targeted detection programme.

Targeted detection programmes are recommended to identify subjects affected by severe alpha1-antitrypsin deficiency (AATD). Guidelines are available to address physicians towards subjects at high risk for AATD. We wanted to investigate the clinical characteristics of subjects enrolled in the programme, who result as not being affected by severe AATD; this information is not available in the present literature. We elaborated data contained in the questionnaires accompanying the samples of 2127 Italian subjects submitted for AATD detection in a period spanning 11 years (1996-2006). A total of 588 subjects were eligible to enter this study: PI*MM subjects and subjects with intermediate AATD, referred for lung disease, were characterised by a relatively young mean age, and a high proportion (31.2% and 28.6%, respectively) were never smokers. Fifty percent or more had symptoms of chronic bronchitis, but without obstruction. Only a minority belonged to most severe GOLD stages. The mean levels of AAT varied as a function of the presence or absence of airflow obstruction in intermediate AATD subjects, but not in PI*MM. Individuals enrolled in AATD detection programmes represent an interesting cohort both for public health and research purposes.

Inflammation and atherosclerosis: the role of TNF and TNF receptors polymorphisms in coronary artery disease.

Inflammation plays an important role in the pathogenesis of atherosclerosis and coronary syndromes; moreover, various lines of evidence suggest that genetic factors contribute significantly to the risk of coronary artery disease (CAD). Through its effects on endothelial function, coagulation, insulin resistance and lipid metabolism, the proinflammatory cytokine TNF could be involved in cardiovascular pathophysiology. The aim of our study is to analyze whether TNF gene promoter (-308 G/A; -857 G/A) and TNF receptor polymorphisms (TNFR1 MspA1 I exon 1 and TNFR2 Nla III exon 6) show involvement in CAD predisposition in a group of Italian patients compared with healthy controls. Genotyping was performed by PCR-RFLP. Consecutive Italian patients with angiographically proven CAD (n= 248) were compared with controls (n=241), matched for age, sex and geographical origins. CAD patients showed a higher frequency of the TNF -308 A allele than healthy controls (p=0.046). After stratification according to risk factors for CAD, our analysis revealed that CAD patients with diabetes (p=0.042) and CAD patients without hypertension (p= 0.0495) displayed a higher frequency of the TNF -308 AA genotype compared with healthy controls. Our data stress the inflammatory nature of CAD and show a possible involvement of TNF -308G/A promoter polymorphisms in the predisposition to the development of this disease. The less frequent A allele seems to be a predisposing factor for development of CAD in particular pathological settings associated with the disease itself, such as diabetes.

Anesthetic management of a combined heart and liver transplantation in an amyloidotic patient: a case report.

Few cases of combined heart and liver transplantation (CHLT) have been reported for familial amyloidosis. Our first CHLT was performed on a female patient with familial amyloidosis due to a genetic defect in transthyretin, characterized by deposition of amyloid in various organs and tissues. This disease produced autonomic heart dysfunction that preceded the development of clinical manifestations and may be an important factor in determining the optimal timing for liver transplantation. CHLT can be performed successfully, even in patients with advanced disease. However, the most compromised patients are more exposed to intraoperative risks, postoperative complications, and worsening of extracardiac and extrahepatic symptoms. Our patient presented severe cardiac dysfunction requiring CHLT. The operative technique is far from being consolidated, despite this, both organs were transplanted in the same day with 2 hours in the intensive care unit (ICU) between surgeries. The outcome of both organs has been favorable. The amyloidotic liver was transplanted to another patient, a sequential (domino) transplantation.

Development of a new protocol for rapid bacterial identification and susceptibility testing directly from urine samples.

The current gold standard method for the diagnosis of urinary tract infections (UTI) is urine culture that requires 18-48 h for the identification of the causative microorganisms and an additional 24 h until the results of antimicrobial susceptibility testing (AST) are available. The aim of this study was to shorten the time of urine sample processing by a combination of flow cytometry for screening and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for bacterial identification followed by AST directly from urine. The study was divided into two parts. During the first part, 675 urine samples were processed by a flow cytometry device and a cut-off value of bacterial count was determined to select samples for direct identification by MALDI-TOF-MS at ≥5 × 10(6) bacteria/mL. During the second part, 163 of 1029 processed samples reached the cut-off value. The sample preparation protocol for direct identification included two centrifugation and two washing steps. Direct AST was performed by the disc diffusion method if a reliable direct identification was obtained. Direct MALDI-TOF-MS identification was performed in 140 urine samples; 125 of the samples were positive by urine culture, 12 were contaminated and 3 were negative. Reliable direct identification was obtained in 108 (86.4%) of the 125 positive samples. AST was performed in 102 identified samples, and the results were fully concordant with the routine method among 83 monomicrobial infections. In conclusion, the turnaround time of the protocol described to diagnose UTI was about 1 h for microbial identification and 18-24 h for AST.

Improved method for the allelic definition of C4A and C4B polymorphism (HLA class III).

The polymorphism of the fourth component of human serum complement (C4) is well established at the proteinic level; at the DNA level in the analysis of C4A and C4B gene polymorphism, the PCR technique is not widely and routinely used because it is time consuming and still presents reproducibility problems. This is a serious problem because only PCR genotyping allows the establishment of Rodgers-Chido reverse antigenicity without the need for classical family segregation studies, whose samples are not always easy to obtain. The most commonly used protocol requires an initial PCR followed by nested amplification of all the products supposed positive or negative. The two reactions are set up using differing cycling conditions, primers, and magnesium chloride concentrations. We developed a simplified procedure to easily obtain reproducible results and used a single protocol for all reactions. Nested PCR is made using only the positive samples, so we decrease the number of samples to handle, the time spent for the work, and the reagents used for the reactions. Moreover, we increased the reproducibility of the experiments.

[Intraoperative anesthetic management of kidney failure in adult liver transplantation. Conventional hemodialysis]. / Manejo anestésico intraoperatorio del fallo renal en el trasplante hepático del adulto. Hemodiálisis convencional.

OBJECTIVE: To evaluate our application of indications, use and benefits of conventional hemodialysis during surgery in patients with advanced liver disease and acute or chronic renal failure undergoing liver transplantation (LP), liver retransplantation (LRT) or combined hepatorenal transplantation (CHRT). PATIENTS AND METHODS: We retrospectively reviewed the cases of 22 patients with advanced liver disease, 11 with acute renal failure and 11 with chronic renal failure. We performed 6 LT, 5 LRT and 11 CHRT. The following data were recorded in the periods before, during and immediately after surgery: metabolic, hemodynamic and coagulation parameters; bicarbonate, calcium and inotropic drug requirements; incidences during reperfusion of the graft; surgical technique used; and survival. RESULTS: Seven patients (32%) needed hemodialysis, 4 (18%) needed ultrafiltration, 7 (32%) needed both and 4 (18%) required neither. For 6 patients total clamping of the inferior vena cava (ICV) was required with external venovenous bypass. For 8 patients total clamping of the IVC was performed without venovenous bypass. For 8 others IVC clamping was partial with retrohepatic preservation (piggy-back). There were 2 deaths during surgery, 4 more within the first month after surgery and 4 more in the second month. Overall survival was 36.4% among acute patients and 72.7% among CHRT patients. CONCLUSIONS: 1) Conventional hemodialysis during surgery is feasible and gives good results; 2) conventional "high efficiency" hemodialysis is more effective and useful in these patients than is either slow, continuous hemodialysis or filtration; 3) the survival rate of CHRT patients is similar to that of patients undergoing LT with normal kidney function, and 4) partial IVC clamping in the anhepatic phase may decrease the need for ultrafiltration.

[Cruorifilaria tuberocauda (Nematoda: Filarioidea) in Venezuelan capybaras]. / La Cruorifilaria tuberocauda (Nematoda: Filarioidea) en capibaras de Venezuela.

Four filariae are known to parasitize the capybara "Hydrochoerus hydrochaeris" namely the "Cruorifilaria tuberocauda", Eberhard, Morales & Orihel (1976). The "Dipetalonema (Alafilaria) hydrochoerus", Yates (1983), and two "mansonella" recently described by Eberhard, Campo-Aasen & Orihel (1983), "Mansonella longicapita" and "Mansonella rotundicapita". In this study we describe the adult parasites and the microfilaria of the "Cruorifilaria tuberocauda" (Nematoda, Filarioidea), Eberhard, Morales & Orihel (1976) obtained from the arteries of the kidney of two capybaras from Apure and Barinas states of Venezuela. The pathology produced by the presence of the filariae in kidney and lungs of the animals is described as well as the blood harboring microfilaria.

Extended spectrum ß-lactamase-producing Escherichia coli faecal carriage in Spanish travellers returning from tropical and subtropical countries.

The aim of this study was to investigate the prevalence of extended-spectrum ß-lactamase (ESBL) -producing Escherichia coli in stool samples from 457 patients with travellers' diarrhoea who had travelled to tropical and subtropical countries. Ninety-seven ESBL-producing E. coli strains were isolated from 17.9% of the patients (82/457). CTX-M-15 was the most prevalent enzyme (80%) and India was the most visited country and showed the highest prevalence of positive samples (37.4%).

Comparison of active and passive adhering leads for endocardial pacing.

The frequency of lead failure requiring invasive correction in a total of 276 implants of four different transvenous leads (6907, continuous lead, IE-65-I, and MIP 2000) was observed during a one-and-one-half year period with a minimum of two months follow-up post-implant. Implants were on a successive sequential basis, randomly distributed between the two surgeons normally performing implants, and unselected for presumed ease or difficulty of the procedure. Failure rates with the 6907 and continuous leads were 7 of 76, or 9.2%; with the IE-65-I, 2 of 76, or 2.6%; and with the MIP 2000, 8 of 45, or 17.8%. The difference between the IE-65-I and the two conventional leads was significant at the 5% level, and between the IE-65-I and the group of the other three at the 1.6% level. The MIP 2000 was significantly different from the other three leads at the 2.7% level. Previous clinical experience with 849 implants with continuous and 6807 leads indicated that the overall data was similar to that obtained in the present evaluation. No significant differences in failure rates between surgeons and no measurable "practice effect" could be detected. It was concluded that the design of the lead is a major factor in the differing need for early secondary intervention.

Comparison for active and passive leads for endocardial pacing--II.

Four commercially available transvenous pacing leads were evaluated in a series of 240 implants on a rotational order basis. Total (intra- and post-operative) failure rates for this series were: Cordis 1 mm, 1/60 (1.7%); Biotronik IE-65-I, 5/60 (8.3%); Medtronic 6961, 4/60 (6.7%); and CPI 4116, 6/60 (10.0%). Post-operative failure rates were: 1 mm, 1/60 (1.7%); IE-65-I, 4/59 (6.8%); 6961, 0/56 (0.0%), and 4116, 3/57 (5.3%). In a previous study conducted at this center with identical protocol, total failure rates were: Medtronic 6907, 7/76 (9.2%); Cordis CL, 7/76 (9.2%); Biotronik IE-65-I, 2/76 (2.6%); and the Vitatron MIP-2000, 9/45 (20.0%). Post-operative failure rates were: 6907, 4/73 (5.5%); CL, 6/75 (8.0%); IE-65-I, 2/76 (2.6%); and MIP-2000, 5/41 (12.2%). If the two IE-65-I series are combined, the total failure rate is 7/136 (5.1%), and the post-operative failure rate is 6/135 (4.4%). Significant differences exist between the highest and lowest failure rates in total and post-operative cases for each series. The final decision concerning use of a particular lead must, of course, be based on clinical criteria.