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Neutrophils have been thought to play a critical role in terminal differentiation of NK cells. Whether this effect is direct or a consequence of global immune changes with effects on NK-cell homeostasis remains unknown. In this study, we used high-resolution flow and mass cytometry to examine NK-cell repertoires in 64 patients with neutropenia and 27 healthy age- and sex-matched donors. A subgroup of patients with chronic neutropenia showed severely disrupted NK-cell homeostasis manifesting as increased frequencies of CD56bright NK cells and a lack of mature CD56dim NK cells. These immature NK-cell repertoires were characterized by expression of the proliferation/exhaustion markers Ki-67, Tim-3, and TIGIT and displayed blunted tumor target cell responses. Systems-level immune mapping revealed that the changes in immunophenotypes were confined to NK cells, leaving T-cell differentiation intact. RNA sequencing of NK cells from these patients showed upregulation of a network of genes, including TNFSF9, CENPF, MKI67, and TOP2A, associated with apoptosis and the cell cycle, but different from the conventional CD56bright signatures. Profiling of 249 plasma proteins showed a coordinated enrichment of pathways related to apoptosis and cell turnover, which correlated with immature NK-cell repertoires. Notably, most of these patients exhibited severe-grade neutropenia, suggesting that the profoundly altered NK-cell homeostasis was connected to the severity of their underlying etiology. Hence, although our data suggest that neutrophils are dispensable for NK-cell development and differentiation, some patients displayed a specific gap in the NK repertoire, associated with poor cytotoxic function and more severe disease manifestations., (© 2022 by The American Society of Hematology.)

Cases of Legionnaires' disease associated with biological treatment plants (BTPs) have been reported in six countries between 1997 and 2010. However, knowledge about the occurrence of Legionella in BTPs is scarce. Hence, we undertook a qualitative and quantitative screening for Legionella in BTPs treating waste water from municipalities and industries in Norway, to assess the transmission potential of Legionella from these installations. Thirty-three plants from different industries were sampled four times within 1 year. By cultivation, 21 (16%) of 130 analyses were positive for Legionella species and 12 (9%) of 130 analyses were positive for Legionella pneumophila. By quantitative real-time polymerase chain reaction (PCR), 433 (99%) of 437 analyses were positive for Legionella species and 218 (46%) of 470 analyses were positive for L. pneumophila. This survey indicates that PCR could be the preferable method for detection of Legionella in samples from BTPs. Sequence types of L. pneumophila associated with outbreaks in Norway were not identified from the BTPs. We showed that a waste water treatment plant with an aeration basin can produce high concentrations of Legionella. Therefore, these plants should be considered as a possible source of community-acquired Legionella infections.

Introduction: This is a follow up study on rescue workers participating in the primary rescue during and immediately after the explosion of a firework factory. We aimed to estimate the possible PTSD prevalence at five and 18 months post disaster, determining if the level of PTSD symptoms at 18 months could be predicted from factors measured at five months., Methods: We included measures of posttraumatic symptoms, social support, locus of control and demographic questions., Results: The possible PTSD prevalence rose from 1.6% (n = 465) at five months post disaster to 3.1% (n = 130) at 18 months. A hierarchical linear regression predicted 59% of PTSD symptoms variance at 18 months post disaster. In the final regression, somatization explained the greatest part of the symptom variance (42%), followed by locus of control (29%) and major life events prior to and right after the disaster (23%)., Conclusion: Rescue workers seemed to be relatively robust to traumatic exposure: The prevalence of possible PTSD in our study was even lower than previous studies, probably because of the less severe consequences of the disaster studied. Furthermore, we found that PTSD symptom level at 18 months post disaster was highly predicted by psychological factors, particularly by somatization. However, further investigations of traumatic responding are required in this population.

Background: The first instance of Legionella infection in a Norwegian hospital was confirmed in 2005. We describe the best-known methods of eradicating Legionella in hospitals., Materials and Method: The article is based on the authors' experience of measures to prevent Legionnaires' disease in hospitals and on a non-systematic search in PubMed., Results: There are several methods of combating Legionella in hospitals. These include chlorination, heat treatment, and the use of filters. However, recontamination easily re-occurs after eradication. The silver and copper ionisation treatment of water is a well-documented method for the systematic and long-term eradication of Legionella in water. The disadvantages of this method are that it is expensive, that there is a risk of discolouring the water, and that there is a possibility of developing resistance in environmental bacteria. This resistance mechanism can theoretically be transferred to bacteria that cause illness., Interpretation: We recommend the silver and copper ionisation treatment of water as a method of preventing nosocomial Legionnaires' disease when standard methods fail and there is a high prevalence of Legionella in the water. The discolouration of operation instruments that occurs as a result of high silver concentrations can be avoided by using a separate water supply for operation units.

Three outbreaks of Legionnaires' disease were reported in the Fredrikstad/Sarpsborg community, Norway, in 2005 and 2008 caused by the L. pneumophila ST15 and ST462 strains determined by sequence based typing. In this retrospective study, we suggest that the aeration ponds, a part of the biological treatment plant at Borregaard Ind. Ltd., are the main amplifiers and primary disseminators of the outbreak L. pneumophila strains. This result is supported by the finding that the ST15 and ST462 strains were not able to survive in air scrubber liquid media more than two days of incubation at the scrubber's operating conditions during the 2005 and 2008 outbreaks. In 2008, >10¹⁰ CFU/L of L. pneumophila ST462 were detected in the aeration ponds. ST15 and ST462 were also detected in the river Glomma in 2005 and 2008, respectively, downstream of the wastewater outlet from the treatment plant (10⁵CFU/L). These findings strongly suggest that the presence of L. pneumophila in the river is due to the release of wastewater from the industrial aeration ponds, demonstrating that the river Glomma may be an additional disseminator of L. pneumophila during the outbreaks. This work emphasizes the need for preventive actions against the release of wastewater containing human pathogens to the environment.

Biological treatment plants are frequently used to degrade organic substances in wastewater from wood refinement processes. Aeration ponds in such plants provide an optimal growth environment for many microorganisms, including Legionella species. To investigate whether legionellae could be dispersed as aerosols from the ponds and transported by the wind, the wetted-wall cyclone SASS 2000(PLUS) and the impactors MAS-100 and STA-204 were used to collect air samples directly above, upwind, and downwind of aeration ponds during a 4-month period. Computational fluid dynamics was used a priori to estimate the aerosol paths and to determine suitable air-sampling locations. Several Legionella species, including Legionella pneumophila, were identified in air samples at the biological treatment plant using microbiological and molecular methods. L. pneumophila was identified up to distances of 200 m downwind from the ponds, but, in general, not upwind nor outside the predicted aerosol paths. The highest concentration level of viable legionellae was identified directly above the aeration ponds (3300 CFU/m3). This level decreased as the distance from the aeration ponds increased. Molecular typing indicated that a single clone of L. pneumophila was dispersed from the ponds during the period of the study. Thus, our study demonstrated that aerosols generated at aeration ponds of biological treatment facilities may contain L. pneumophila, which then can be transported by the wind to the surroundings. The methods used in this study may be generically applied to trace biological aerosols that may pose a challenge to environmental occupational health.

Background: On 21 May 2005, the Norwegian health authorities were alerted by officials from a local hospital that several recent patients had received the diagnosis of legionnaires disease; all patients resided in 2 neighboring municipalities. We investigated the outbreak to identify the source and to implement control measures., Methods: We interviewed all surviving case patients and investigated and harvested samples from 23 businesses with cooling towers and other potential infection sources. The locations of the businesses and the patients' residences and movements were mapped. We calculated attack rates and risk ratios among people living within various radii of each potential source. Isolates of Legionella pneumophila were compared using molecular methods., Results: Among 56 case patients, 10 died. The case patients became ill 12-25 May, resided up to 20 km apart, and had not visited places in common. Those living up to 1 km from a particular air scrubber had the highest risk ratio, and only for this source did the risk ratio decrease as the radius widened. Genetically identical L. pneumophila serogroup 1 isolates were recovered from patients and the air scrubber. The air scrubber is an industrial pollution-control device that cleans air for dust particles by spraying with water. The circulating water had a high organic content, pH of 8-9, and temperature of 40 degrees C. The air was expelled at 20 m/s and contained a high amount of aerosolized water., Conclusions: The high velocity, large drift, and high humidity in the air scrubber may have contributed to the wide spread of Legionella species, probably for >10 km. The risk of Legionella spread from air scrubbers should be assessed.

Polymerase chain reaction (PCR) and Southern blotting were used to assess the prevalence of HPV in cervical biopsies of 100 women who were treated at the gynecology department of Telemark Central Hospital for non-cancerous conditions. Nine (9%) of the biopsies were HPV positive. Four (4%) were of HPV type 18, one (1%) was HPV11 positive, and four contained different unrecognized HPV types (HPVX). HPV16 was not detected.

We tested two sets of primers derived from the dhps gene of Neisseria meningitidis for the amplification of meningococcal DNA by PCR. Both the NM1-NM6 primers and the NM3-NM6 primers amplified dhps DNA from all of the meningococci included in the study, resulting, in most cases, in amplicons of 0.70 and 0.23 kb, respectively. Also, dhps DNAs of N. gonorrhoeae and some commensals were amplified but Haemophilus influenzae, Streptococcus pneumoniae, and Escherichia coli DNAs were not. By PCR amplicon restriction endonuclease analysis (AREA) of the larger amplicon, we could differentiate between individual strains of N. meningitidis. Following two cases of meningococcal disease, we used PCR AREA to identify healthy contacts carrying the disease-causing strain. We conclude that PCR AREA is a useful method for meningococcal strain differentiation and that it has potential as a method for studying the spread of a disease-causing strain in an affected population. The method is quicker and easier to perform and interpret than chromosomal DNA fingerprinting.

Chromosomal DNA fingerprinting indicated that Norwegian Taylorella equigenitalis strains are genetically homogeneous and similar to some Swedish isolates but different from other European strains. As contagious equine metritis is rarely a serious disease in Norwegian horses, we conclude that the dominant T. equigenitalis strain in Norway is a genetically homogeneous clone of low virulence.

The objective of the present study was to determine the prevalence of human papillomavirus (HPV) infections in Norwegian women with cervical cancer. We used the polymerase chain reaction (PCR) and Southern blot techniques to assess the prevalence of HPV in cervical biopsies of 133 women admitted to the Norwegian Radium Hospital for treatment of cervical cancer. At the time of sampling (from February 1988 to April 1989) about 85% of Norwegian women with cervical cancer were treated at the Norwegian Radium Hospital. HPV was found in biopsies of 91 (68%) of women with cancer; 70 (53%) biopsies contained HPV type 16, 19 (14%) HPV type 18, 4 (3%) HPV type 33, 2 (1.5%) HPV type 11, and 3 (2%) HPV DNA of unknown type (HPVX). Five percent of biopsies were doubly infected, chiefly with HPV 16 + 18. We found a significant association between HPV 18 and low age, poorly differentiated tumors and adenocarcinomas. Our results show that there is an association between HPV types 16 and 18 and cervical cancer also in a Norwegian setting. PCR was more sensitive than Southern blotting for detection of HPV. Thirty-six (27.5%) of cancer biopsies were positive by PCR but negative by Southern blotting, as against 49 (73.5%) positive by both methods; we also encountered 4 samples positive by Southern blotting and negative by PCR. In 23/53 cancer biopsies positive by Southern blotting we found evidence for integrated or rearranged HPV genomes.

Since 1987 we have analysed throat samples from 1,086 healthy contacts of 32 patients with meningococcal disease. The disease-causing strain was found in contacts of 17 out of the 32 patients. 161 (18%) of the contacts carried meningococci, and 30 (3%) of them were carriers of the disease-causing strain as determined by DNA fingerprinting. The carrier strain was eradicated in 29 of these 30 contacts by treatment with rifampicin. No secondary case of meningococcal disease was observed. During the four-year period 1984-87, there were 39 confirmed cases of meningococcal disease, including 12 verified and four suspected secondary cases of meningococcal disease. Therefore identification and eradication of the disease-causing strain seems to prevent secondary cases. A change in the Norwegian recommendations for preventing secondary cases of meningococcal disease should be discussed.

PCR was used to detect HPV DNA in cervical scrapes of 23 women with cervical cancer. Compared with PCR-assisted HPV DNA detection in parallel biopsies, the sensitivity was 81% and the specificity 100%. We conclude that cervical scrapes can be used for detection of HPV DNA in women with cervical cancer.

In Norway, the use of chemoprophylaxis after cases of meningococcal disease is not recommended. Instead, household members less than 15 years are treated with penicillin for 7 days. Failures of this treatment have been reported. We therefore used DNA fingerprinting to identify the disease-causing strain in healthy contacts combined with selective rifampicin prophylaxis to these carriers to prevent secondary cases. During a 2-year period (1987-89) there were 13 cases of meningococcal disease in the County of Telemark (165000 inhabitants). 65 (14.7%) out of 441 contacts to these 13 patients harbored meningococci in their throat; 16 (3.6%) carried the disease-causing strain. Only 1 carrier fulfilled the criteria for being treated with penicillin; 8 were adults and the remaining 7 were not household members. No secondary cases of meningococcal disease occurred during the study period or the following 12 months. During the 4-year period (1984-87) preceding the study period there were 39 cases of meningococcal disease in Telemark; 7 of them were index cases for 12 bacteriologically verified and 4 clinically suspected secondary cases of meningococcal disease. We conclude that selective prophylaxis with rifampicin seems to be more efficient that penicillin treatment of household members less than 15 to prevent secondary cases of meningococcal disease.

We describe the detection of eight genital human papillomavirus (HPV) types, including HPV16 and HPV18, by PCR amplification of a 323 base-pair region of the genome within the L1 open reading frame (ORF). The primer sequences are: TGYAAATATCCWGATTWTWT and GTATCWACMACAGTAACAAA. The method will detect purified HPV16 DNA down to a concentration of as little as a single molecule in 100 microliters. The method is also applicable to purified DNA and crude lysates from tumour biopsies. Typing of the PCR product can be achieved with specific oligonucleotide probes.

Rapid diagnosis of meningococcal disease followed by early treatment is essential. However, culture of blood or cerebrospinal fluid (CSF) may be unsuccessful because antibiotic treatment is often started before adequate specimens are collected, and because bacteria may die during transportation to the laboratory. We have used the polymerase chain reaction (PCR) to detect meningococcal DNA in a culture-negative CSF of a 15-year-old girl with meningococcal disease. Two oligonucleotides flanking the dihydropteroate synthase gene (dhps) of Neisseria meningitidis were used as primers. The PCR reaction is a rapid technique for the early detection of meningococcal meningitis, and also when culture is negative.

During a study on the epidemiology of Staphylococcus aureus colonization in newborns, mothers, and hospital staff, S. aureus was isolated from 536 of 1,945 specimens. Ninety-three isolates of S. aureus from the three groups of individuals were included in a study to evaluate the potential of DNA fingerprinting for strain differentiation. The 93 isolates were also phage typed and their plasmid profiles were analyzed. Cleavage of DNA with BamHI resulted in 13 different DNA restriction endonuclease band patterns (DNA REBPs), one of which consisted of eight isolates whose DNA was not cleaved with BamHI. The DNAs from these eight isolates were easily cleaved with HindIII. The different DNA REBPs were stable both during in vitro and in vivo growth and allowed strain differentiation within phage groups or types. We could not show any strong association between DNA REBP classes, phage types or groups, and plasmid profiles. Of the 93 isolates, 27 (29.0%) could not be phage typed and 12 (12.9%) lacked plasmids. We therefore conclude that DNA fingerprinting is a powerful tool, in addition to phage typing and plasmid profile analysis, for strain differentiation of S. aureus.

By cloning studies and complementation experiments, the sulfonamide resistance gene of a serogroup B and serotype 15 (B:15) strain of Neisseria meningitidis was localized to a 1.2-kb chromosomal SspI fragment expressing a drug-resistant dihydropteroate synthase. The fragment hybridized to DNA from both resistant and susceptible strains, suggesting that the resistance gene is a variant of the normal gene for dihydropteroate synthase.

During the period from January 1987 to June 1988 DNA fingerprinting was used to identify carriers of the disease-causing strain of Neisseria meningitidis among the patients contacts. A total of 432 persons were screened during nine episodes. The overall carrier-rate was 16.2%, and the causative strain was found in 3.4% of the contacts. Eleven carriers were successfully treated with rifampicin, whereas two of three carriers treated with penicillin remained carriers. We conclude that DNA fingerprinting is a valuable tool for rapid identification of carriers of the causative organism in order to eradicate the epidemic strain of N. meningitidis.

Plasmid profile analysis, restriction endonuclease analysis, and multilocus enzyme electrophoresis were used in conjunction with serotyping, bacteriophage typing, and biochemical fingerprinting to trace epidemiologically related isolates of Salmonella typhimurium from an outbreak caused by contaminated chocolate products in Norway and Finland. To evaluate the efficiency of the epidemiological marker methods, isolates from the outbreak were compared with five groups of control isolates not known to be associated with the outbreak. Both plasmid profile analysis and phage typing provided further discrimination over that produced by serotyping and biochemical fingerprinting. Plasmid profile analysis and phage typing were equally reliable in differentiating the outbreak isolates from the epidemiologically unrelated controls and were significantly more effective than multilocus enzyme electrophoresis and restriction enzyme analysis of total DNA. The greatest differentiation was achieved when plasmid profile analysis and phage typing were combined to complement serotyping and biochemical fingerprinting. However, none of the methods employed, including restriction enzyme analysis of plasmid DNA, were able to distinguish the outbreak isolates from five isolates recovered in Norway and Finland over a period of years from dead passerine birds and a calf.