Changes in the Quality and Microbial Communities of Precooked Seasoned Crayfish Tail Treated with Microwave and Biological Preservatives during Room Temperature Storage.

The qualities of precooked foods can be significantly changed by the microorganisms produced during room temperature storage. This work assessed the effects of different antibacterial treatments (CK, without any treatment; microwave treatment, MS; microwave treatment and biological preservatives, MSBP) on the physicochemical properties and microbial communities of precooked crayfish tails during room temperature storage. Only the combination of microwave sterilization and biological preservatives significantly inhibited spoilage, as evidenced by the total viable count (4.15 log CFU/g) after 3 days of room temperature storage, which satisfied the transit time of most logistics companies in China. Changes in pH and TVB-N were also significantly inhibited in the MSBP group compared with those in the CK and MS groups. More than 30 new volatile compounds were produced in the CK groups during room temperature storage. However, in the MSBP groups, the volatile compounds were almost unchanged. The correlations between the microbial composition and volatile compounds suggested that specific bacterial species with metabolic activities related to amino acid, energy, cofactor, and vitamin metabolism, as well as xenobiotics biodegradation and metabolism, were responsible for the changes in volatile compounds. These bacteria included Psychrobacter, Arthrobacter, Facklamia, Leucobacter, Corynebacterium, Erysipelothrix, Devosia, Dietzia, and Acidovorax. Overall, our findings provide a foundation for the development of strategies to inhibit spoilage in precooked crayfish tails stored at room temperature.

Klebsiella pneumoniae invasive syndrome with pneumocephalus and extensive cerebral infarction: Case report.

A 54-year-old female with diabetes was admitted with fever and altered consciousness. Laboratory tests revealed venous blood glucose level of 43.79 mmol/L. Computed tomography (CT) scans of the head, chest, and abdomen showed a right-sided pneumothorax, consolidation, and atelectasis in the right lung; a large heterogeneous density lesion with fluid and gas-fluid levels in the liver; and scattered gas shadows in both kidneys, respectively. Blood and puncture fluid cultures indicated infection with Klebsiella pneumoniae. Based on the susceptibility profiles of the isolates, imipenem was administered intravenously to treat the infection. On the third day of hospitalization, the patient's condition worsened, with head CT showing an extensive cerebral infarction and multiple gas accumulations in the right cerebral hemisphere, as well as a large-area cerebral infarction in the left parietal and occipital lobes. Ultimately, the patient died of multiple organ dysfunction on the fourth day after initial presentation. Although the Klebsiella pneumoniae isolates from the patient showed sensitivity to imipenem, this antibiotic shows poor entry into the central nervous system. The death of the patient indicates that the selection of antibiotics that can cross the blood-brain barrier may be crucial in the outcome of this type of case. Therefore, antibiotics that can penetrate the blood-brain barrier should be selected as soon as possible, and empirical treatment must be initiated immediately after clinical suspicion of invasive Klebsiella pneumoniae, even if the diagnosis has not been determined.

Case report: Trigeminal neuralgia misdiagnosed as glossopharyngeal neuralgia.

Background: Trigeminal neuralgia (TN) and glossopharyngeal neuralgia (GPN) are cranial nerve neuralgias with the same clinical manifestations, pathological features, and trigger factors; their affected sites are adjacent. Performing a magnetic resonance imaging (MRI) examination alone can easily lead to a misdiagnosis. Case presentation: A 72-year-old man had visited another hospital with severe left-sided tongue pain. On MRI, vascular compression of the glossopharyngeal nerve had been visible, with unclear evidence of trigeminal nerve involvement. He had been diagnosed with left-sided GPN and underwent microvascular decompression (MVD) of the left glossopharyngeal nerve. However, no improvement was observed after surgery. During a second surgery at our hospital, MVD of the trigeminal nerve was performed, and the trigeminal nerve was fully explored and separated. The patient's pain resolved after surgery. Ultimately, the patient was definitively diagnosed with left-sided TN. Discussion and conclusion: MVD is currently the most efficacious surgical option for treating cranial nerve neuralgia. To select patients for MVD, having an MRI criteria for identifying true neurovascular compression will be helpful. However, clinicians should focus more on a patient's clinical symptoms and not rely solely on MRI findings. This patient's case can help clinicians distinguish between TN and GPN, improve the understanding of these diseases, avoid misdiagnosis, and reduce the possibility of secondary damage.

Detection of co-infection with Orientia tsutsugamushand and hemorrhagic fever with renal syndrome by next-generation sequencing.

Purpose: The co-infection with Orientia tsutsugamushand and hemorrhagic fever with renal syndrome is rare. There are many similarities in early clinical practice between the two diseases, and sometimes it is easy to misdiagnose, especially when co-infection occurs.Methods: We describe a patient who presented with fever and headache after bitten by an insect and whose physical examination showed conjunctival hyperemia, eschar and petechiae in tongue and the soft palate. To lead to a diagnosis, the serum antibody of Hantaan virus, Weil-Felix test and  next-generation sequencing of cerebrospinal fluid was performed.Results: The Weil-Felix test was negative on the 15th day after the onset of the disease and a repeated Weil-Felix test on the 21st day showed a titer of 1:160 and the IgM against Hantaan virus was positive. The number of sequence reads identified corresponding to O. tsutsugamushi was 239 with a genomic coverage of 0.9178%. This patient was diagnosed with intracranial infection with Orientia tsutsugamushi and co-infection with epidemic hemorrhagic fever. The symptoms in our patient quickly decreased after the administration of tetracycline.Conclusion: Next-generation sequencing is helpful for the early diagnosis of scrub typhus, especially when the Weil-Felix test is negative. Clinicians need to be reminded to screen for common pathogens that may be co-infected, such as epidemic hemorrhagic fever.

Associations between plasma metal levels and mild renal impairment in the general population of Southern China.

BACKGROUND: Metal exposure were assumed to be closely related with declined renal function, but the conclusions were controversial. We employed diverse statistical models and assessed the association between metal mixture exposure and mild renal impairment. METHODS: A total of 13 plasma metals were measured in 896 general population from Southern China. Subjects with estimated glomerular filtration rate within 60-89 ml/min/1.73 m2 and urinary albumin-creatinine ratio <30 mg/g creatinine were defined as mild renal impairment (MRI). RESULTS: About 31.47 % participants showed MRI. In the multivariate logistic regression models, compared with the first quartile, high levels of arsenic and molybdenum (the fourth quartile) were both associated with MRI, and the ORs (95 % CI) were 1.68 (1.05, 2.68) and 2.21 (1.40, 3.48), respectively. Their predominant roles were identified by the weighted quantile regression (WQS). Besides, restricted cubic spline analysis verified the relationship between molybdenum level and increased MRI risk in a linear and dose-response manner. CONCLUSION: High levels of arsenic and molybdenum might be independent risk factors of MRI, and they showed combined effect. Our findings might provide vigorous evidence in preventing mild decline in renal function.

Central Hypoventilation Is a Key Risk Factor for Mechanical Ventilation During the Acute Phase of Anti-N-Methyl-D-Aspartate Receptor Encephalitis.

Objective: Anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis is an acute form of encephalitis of autoimmune etiology. We aimed to evaluate the risk factors that predicted the need for mechanical ventilation during the acute phase of anti-NMDAR encephalitis through an analysis of the clinical characteristics and biochemical test results of the patients with anti-NMDAR encephalitis. Methods: In this retrospective study, patients who primarily presented with anti-NMDAR encephalitis and exhibited anti-NMDAR antibody positivity in the cerebrospinal fluid (CSF) between November 2015 and February 2020 were included. Data on the clinical characteristics, biochemical test results, and treatment methods selected for the patients were collected for the analysis of factors predicting the need for mechanical ventilation. Results: Thirty-one patients with a median age of onset of 31 years (inter-quartile range: 21-48 years) were included in this study, of which 15 were male (48.4%). Psychosis (23, 74.2%), seizures (20, 64.5%), and memory deficit (20, 64.5%) were the most common clinical manifestations. At admission, 17 patients (54.8%) presented with pyrexia, of which 12 (38.7%) had a body temperature ≥38°C, and six patients (19.4%) presented with central hypoventilation. All patients received first-line therapy (glucocorticoids, intravenous immunoglobulin, or plasmapheresis alone or combined), whereas two patients (6.5%) received rituximab, a second-line agent, as well. Seven patents required mechanical ventilation. Results of univariate logistic regression analysis revealed that body temperature ≥38°C [odds ratio (OR) = 18, 95% confidence interval (CI): 1.79-181.31, P < 0.05] and central hypoventilation at admission (OR = 57.50, 95% CI: 4.32-764.89, P < 0.05) were the risk factors for mechanical ventilation. Multivariate logistic regression analysis showed that central hypoventilation at admission was the only risk factor predicting the need for mechanical ventilation. Conclusion: Central hypoventilation at admission is a key risk factor for mechanical ventilation during hospitalization in patients with anti-NMDAR encephalitis.

Characteristics of Anti-Contactin1 Antibody-Associated Autoimmune Nodopathies With Concomitant Membranous Nephropathy.

Background: The concurrence of anti-contactin 1 (CNTN1) antibody-associated chronic inflammatory demyelinating polyneuropathy (CIDP) and membranous nephropathy (MN) has previously been reported in the literature. CIDP with autoantibodies against paranodal proteins are defined as autoimmune nodopathies (AN) in the latest research. In view of the unclear relationship between CIDP and MN, we performed a case study and literature review to investigate the clinical characteristics of anti-CNTN antibody-associated AN with MN. Methods: We detected antibodies against NF155, NF186, CNTN1, CNTN2, CASPR1 and PLA2R in blood samples of a patient with clinically manifested MN and concomitant peripheral neuropathy via double immunofluorescence staining and conducted a quantitative measurement of anti-PLA2R IgG antibodies via enzyme-linked immunosorbent assay (ELISA). Case reports of anti-CNTN1 antibody-associated AN, anti-CNTN1 antibody-associated AN with MN, and CIDP with MN were retrieved through a literature search for a comparative analysis of clinical characteristics. The cases were grouped according to the chronological order of CIDP and MN onset for the comparison of clinical characteristics. Results: A 57-year-old man with anti-PLA2R positive MN was admitted to the hospital due to limb numbness, weakness, and proprioceptive sensory disorder. He was diagnosed with anti-CNTN1 antibody-associated AN and recovered well after immunotherapy. Our literature search returned 22 cases of CIDP with MN that occurred before, after, or concurrently with CIDP. Good responses were achieved with early single-agent or combination immunotherapy, but eight out of the 22 patients with CIDP and concomitant MN ultimately developed different motor sequelae. Five patients had anti-CNTN1 antibody-associated AN with MN. Among these patients, males accounted for the majority of cases (male:female=4:1), the mean age at onset was late (60.2 ± 15.7 years, range 43-78 years), and 40% had acute to subacute onset. Clinical manifestations included sensory-motor neuropathy, sensory ataxia caused by proprioceptive impairment, and elevated cerebrospinal fluid protein levels. Conclusion: The age at onset of CIDP with MN was earlier than that of anti-CNTN1 antibody-associated AN. MN may occur before, after or concurrently with CIDP. The early detection and isotyping of anti-CNTN1 and anti-PLA2R antibodies and the monitoring of isotype switching may be essential for suspected CIDP patients.

Clinical features, treatments, and outcomes of patients with anti-N-methyl-D-aspartate encephalitis-a single-center, retrospective analysis in China.

Objective: To describe the clinical features, laboratory data, treatment, and outcomes of anti-N-methyl-D-aspartate (NMDAR) encephalitis in Chinese patients. Methods: This retrospective study included hospitalized patients definitively diagnosed with anti-NMDAR encephalitis and positive for anti-NMDAR antibodies in the cerebrospinal fluid (CSF) in Shenzhen People's hospital, between November 2015 and February 2020. The clinical manifestation, laboratory data, treatments and outcomes were collected retrospectively. Patients were followed up for more than 1 year. Results: The study included 31 patients (15 men, 48.4%) with a median age of 31 years (interquartile range 21-48). The most common clinical presentations were psychosis (n = 23, 74.2%), seizures (n = 20, 64.5%), and memory impairment (n = 20, 64.5%). Total magnetic resonance imaging abnormalities were found in 11 patients (35.5%), with the medial temporal and frontal lobes as the most commonly involved. Abnormal electroencephalogram was observed in 16 patients (51.6%). Five out of 31 patients (19.5%) were diagnosed as neoplasm, including five females with ovarian teratoma and one male with a central nervous system tumor. Multiple immune antibodies, including anti-SSA antibody in four patients (15.4%), anti-Ro52 antibody in four (15.4%), antinuclear antibody (ANT) in four (15.4%), anti-thyroglobulin antibodies (TGAb) in five (17.2%), and thyroid peroxidase antibodies (TPOAb) in three (10.3%) were present. All patients received first-line immunization therapy (intravenous immunoglobulin, glucocorticoids, or plasmapheresis alone or combined), and only two patients (7.3%) received second-line immunization therapy (rituximab). Mechanical ventilation was more necessary in women (37.5%) than in men (6.7%) (p = 0.04), and 29 (93.5%) had favorable clinical outcomes. At more than 12 months of follow-up, the median modified Rankin Scale score decreased from 4 to 0. Conclusions: Patients with anti-NMDAR encephalitis in China had high rates of psychosis and seizures, with low rates of underlying neoplasms. A higher proportion of female patients required mechanical ventilation. Complications with other positive autoimmune antibodies were a common clinical symptoms of anti-NMDAR encephalitis. Majority of the patients obtained satisfactory outcomes in combination with early first-line and long-term immunization therapy.

Case Report: Overlapping Multiple Sclerosis With Anti-N-Methyl-D-Aspartate Receptor Encephalitis: A Case Report and Review of Literature.

Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is an autoimmune disorder mediated by NMDAR antibodies, typically manifesting as behavioral complaints, psychosis, seizures, movement disorders, hypoventilation, and autonomic dysfunction. In recent years, the predisposing factors and pathophysiological mechanisms of anti-NMDAR encephalitis have been tried to be clarified. It has been recognized that an overlap may be observed between anti-NMDAR encephalitis and inflammatory demyelinating disease. However, anti-NMDAR encephalitis is rarely associated with multiple sclerosis. Here, we describe a Chinese female patient diagnosed with relapsing remitting multiple sclerosis who developed anti-NMDAR encephalitis. Further, we discuss the previously reported literature.

Blood miR-1275 is associated with risk of ischemic stroke and inhibits macrophage foam cell formation by targeting ApoC2 gene.

Apolipoprotein C2 (ApoC2) is an important member of the apolipoprotein C family and functions as a major activator of lipoprotein lipase (LPL). In cardiovascular and cerebrovascular systems, the lipolytic activity of the LPL-ApoC2 complex is critical for the metabolism of triglyceride-rich lipoproteins and contributes to the pathogenesis of ischemic stroke (IS). However, the regulation of ApoC2 in IS development remains unclear. In this study, we first explored potential ApoC2-targeting microRNAs (miRNAs) by bioinformatics tool and compared the miRNA expression profiles in the blood cells of 25 IS patients and 25 control subjects by miRNA microarray. miR-1275 was predicted to bind with the 3' untranslated region of ApoC2, and a significant reduction of blood miR-1275 levels was observed in IS patients. Dual-luciferase reporter assay and quantitative RT-PCR confirmed the regulation of ApoC2 by miR-1275 in THP-1 derived macrophages. miR-1275 also inhibited cellular uptake of ox-LDL and suppressed formation of macrophage foam cell. Furthermore, the whole blood miR-1275 levels were validated in 279 IS patients and 279 control subjects by TaqMan assay. miR-1275 levels were significantly lower in IS cases and logistic regression analysis showed that miR-1275 level was negatively associated with the occurrence of IS (adjusted OR, 0.76; 95% CI, 0.69-0.85; p < 0.001). Addition of miR-1275 to traditional risk factors showed an additive prediction value for IS. Our study shows that blood miR-1275 levels were negatively associated with the occurrence of IS, and miR-1275 might exert an athero-protective role against the development of IS by targeting ApoC2 and blocking the formation of macrophage foam cells.

Phycoerythrin-phycocyanin aggregates and phycoerythrin aggregates from phycobilisomes of the marine red alga Polysiphonia urceolata.

Phycoerythrin-phycocyanin aggregates and phycoerythrin aggregates showing special spectral characteristics were prepared from the partly dissociated products of phycobilisomes from Polysiphonia urceolata. The absorption difference spectra between the aggregates and phycoerythrins showing normal spectral characteristics show peaks at 583 nm. The fluorescence emission difference spectra between the phycoerythrin aggregates and normal phycoerythrins show peaks at 602 nm. The special spectral characteristics of the PE aggregates disappeared when the PE aggregates were dissociated in deionized water. The intact phycobilisomes show similar characteristics as phycoerythrin-phycocyanin aggregates. When the peak values at 583 nm in absorption difference spectra were compared with those at 615 nm, it can be found that the amount of the 583 nm chromophores in the phycoerythrin-phycocyanin aggregates and intact phycobilisomes is similar to the amount of PCB chromophores in phycocyanins. It can be concluded that the 583 nm chromophores are at the interface between phycoerythrins and phycocyanins in phycobilisomes and transfer the light energy absorbed by other chromophores in phycoerythrins to the PCB chromophores in phycocyanins. A rod linker polypeptide with molecular weight of 40 kDa was found in the phycoerythrin-phycocyanin aggregates and phycoerythrin aggregates, and it is believed to play roles in the spectral red shift of the aggregates.

Off-axis eight-pass neodymium glass laser amplifier with high efficiency and excellent energy stability.

A new relay-imaged off-axial eight-pass laser amplifier with several joules energy and 1 Hz repetition rate was demonstrated. The extraction efficiency and pulse-to-pulse energy stability were greatly improved. Under the single-pass small-signal gain of 3.6, a net gain of 900 and an effective extraction efficiency of 42.4% in the beam aperture were realized. Pulse-to-pulse energy stability of 0.83% (peak-valley) and 0.17% (root-mean-square) was achieved by the significant saturation of eight-pass amplification, which, to the best of our knowledge, is the best energy stability in several-joules-class amplifiers. The far-field quality was 2.52 times the diffraction limit, and the near-field modulation of the 90% beam aperture was 1.28. No parasitic oscillations or pencil beams were observed. Moreover, another key feature of the proposed amplifier was the ability to remarkably improve the pulse contrast with a unique design.

Baseline platelet parameters for predicting early platelet response and clinical outcomes in patients with non-cardioembolic ischemic stroke treated with clopidogrel.

PURPOSE: The present study investigated whether routine baseline platelet parameters(BPPs) detected before clopidogrel therapy in acute non-cardioembolic ischemic stroke(NCIS) could predict early platelet response and future clinical outcomes. RESULTS: The CYP2C19 polymorphisms constituted independent risk factors for LCR. The number of female patients, the incidence of diabetes mellitus (DM), the level of low-density lipoprotein(LDL) cholesterol, and the neutrophil-to-lymphocyte ratio(NLR) were significantly high in the clinical clopidogrel resistance (CCR) group. However, none of the BPPs had a significant association with laboratory clopidogrel resistance (LCR) or discriminated with the cut-off values regarding LCR or CCR. The patients were divided into two groups according to the average mean platelet volume(MPV) or platelet count(PC). We found that the HbA1c level, the number of female patients, and the CCR were higher in the groups with elevated MPV (≥ 10.6fL) and PC (≥ 235 × 109/L); the LCR, the NIHSS score at discharge, and elevated MPV and PC were risk predictors for CCR. MATERIALS AND METHODS: This study included 196 patients with acute NCIS who underwent routine blood tests upon admission, were treated with clopidogrel, and were followed up for 6 months. Early platelet response was assessed and the CYP2C19 genetic variants were screened for. All participants were categorized into either laboratory clopidogrel resistance(LCR) or clinical clopidogrel resistance (CCR) groups. CONCLUSIONS: Elevated baseline MPV and PC before clopidogrel therapy, as well as CYP2C19 gene variants, should be included in a risk algorithm for NCIS. Furthermore, other nongenetic clinical risk factors should be assessed for optimal prediction of the risk for thrombotic events because of individual variability in platelet response to clopidogrel.

Characteristics of an R-phycoerythrin with two γ subunits prepared from red macroalga Polysiphonia urceolata.

An R-phycoerythrin (R-PE) was isolated by gel filtrations on Sepharose CL-4B and Sephadex G-150 from the phycobiliprotein extract of the marine red macroalga Polysiphonia urceolata Grev and further purified by ion exchange chromatography on DEAE-Sepharose Fast Flow. The purified R-PE showed three absorption peaks at 498 nm, 538 nm, 566 nm and one fluorescent emission maximum at 577 nm. Although the R-PE showed a single band on the examination by native PAGE, it exhibited two very close bands at pH about 4.7 in native isoelectric focusing (IEF). Polypeptide analysis of the R-PE demonstrated that it contained four chromophore-carrying subunits, α18.2, ß20.6, γ31.6 (γ'), γ34.6 (γ), and no colorless polypeptide; its subunit composition was 6α18.2:6ß20.6:1 γ31.6:2γ34.6. The α and ß subunits were distributed within a acidic pH range from 5.0 to 6.0 in denaturing IEF and the γ subunits were in a basic pH range from 7.6 to 8.1. These results reveal that the prepared R-PE may exist in two hexamers of γ (αß)3 γ (αß)3γ' and γ (αß)3 γ'(αß)3 γ and that the R-PE participate in the rod domain assembly of P. urceolata phycobilisomes by stacking each of its trimer (αß)3 face-to-face with the aid of one γ subunit (γ or γ').

Phycoerythrins in phycobilisomes from the marine red alga Polysiphonia urceolata.

Phycoerythrins (PE) in phycobilisomes from Polysiphonia urceolata were studied in this research. Dissociative products of phycobilisomes were analyzed by sucrose density gradient centrifugation and native-PAGE. At least three types of PEs were found in the dissociative products of phycobilisomes. According to their molecular weights, absorption spectra and subunit components, they should be PE hexamer containing γ1 subunit, PE hexamer containing γ2 subunit and PE monomer containing no γ subunit. PEs bigger than hexmer were also found in the dissociative products of phycobilisomes in 200mM phosphate buffer when dissociated phycobilisomes were analyzed by sucrose density gradient centrifugation. PE trimers containing no γ subunits were also found in products of dissociated phycobilisomes in deionized water when dissociated phycobilisomes were analyzed by native-PAGE. This is the first time that pure PE hexamers containing γ2 subunits were isolated from P. urceolata. The PE monomers containing no γ subunits should come from PE trimers or hexamers containing no γ subunits in the "rod" of phycobilisomes. It can be concluded that there are three types of PEs in "rod" of phycobilisomes from P. urceolata: PE containing γ1 subunit, PE containing γ2 subunit and PE containing no γ subunit.

Isolation, purification and properties of an R-phycocyanin from the phycobilisomes of a marine red macroalga Polysiphonia urceolata.

Phycobilisomes were prepared from a marine red macroalga Polysiphonia urceolata (P. urceolata) by sucrose step-gradient ultracentrifugation. From the prepared phycobilisomes, an R-phycocyanin was isolated by gel filtration on Sephadex G-150 and then purified by ion exchange chromatography on DEAE-Sepharose Fast Flow and native polyacrylamide gel electrophoresis (PAGE) performed in neutral buffer systems. The purified R-phycocyanins showed not only a homogeneous trimer of 136 kDa in gel filtration and a single band in native PAGE, but also exhibited one band at about pH 5.7 in native isoelectric focusing (IEF). By a gradient SDS-PAGE the purified R-phycocyanin was determined to contain one a subunit of 17.5 kDa (α17.5) and two b subunits of 21.3 kDa and 22.6 kDa (ß21.3 and ß22.6). The analysis from denaturing isoelectric focusing and two-dimension PAGE demonstrated that α17.5, ß21.3 and ß22.6 had their pIs of 6.4, 5.3 and 5.4, respectively. Furthermore, mass spectroscopy analysis of ß21.3 and ß22.6 by MALDI-TOF mass spectrometry demonstrated the two b subunits had differences in peptide mass fingerprinting. These results revealed that the prepared R-phycocyanins were composed of one a and two b subunits. (α17:53 ß21:32 ß22:61) and (α17:53 ß21:31 ß22:62), which have a structural foundation to show their pIs too close for them to be definitely resolved by native IEF, are postulated to be the most possible trimeric forms of the R-phycocyanins prepared from the phycobilisomes of P. urceolata.

Isolation, purification and properties of an R-Phycocyanin from the phycobilisomes of a marine red macroalga Polysiphonia urceolata.

Phycobilisomes were prepared from a marine red macroalga Polysiphonia urceolata (P. urceolata) by sucrose step-gradient ultracentrifugation. From the prepared phycobilisomes, an R-phycocyanin was isolated by gel filtration on Sephadex G-150 and then purified by ion exchange chromatography on DEAE-Sepharose Fast Flow and native polyacrylamide gel electrophoresis (PAGE) performed in neutral buffer systems. The purified R-phycocyanins showed not only a homogeneous trimer of 136 kDa in gel filtration and a single band in native PAGE, but also exhibited one band at about pH 5.7 in native isoelectric focusing (IEF). By a gradient SDS-PAGE the purified R-phycocyanin was determined to contain one α subunit of 17.5 kDa (α (17.5)) and two ß subunits of 21.3 kDa and 22.6 kDa (ß (21.3) and ß (22.6)). The analysis from denaturing isoelectric focusing and two-dimension PAGE demonstrated that α (17.5), ß (21.3) and ß (22.6) had their pIs of 6.4, 5.3 and 5.4, respectively. Furthermore, mass spectroscopy analysis of ß (21.3) and ß (22.6) by MALDI-TOF mass spectrometry demonstrated the two ß subunits had differences in peptide mass fingerprinting. These results revealed that the prepared R-phycocyanins were composed of one α and two ß subunits. and , which have a structural foundation to show their pIs too close for them to be definitely resolved by native IEF, are postulated to be the most possible trimeric forms of the R-phycocyanins prepared from the phycobilisomes of P. urceolata.

The 42.1 and 53.7 kDa bands in SDS-PAGE of R-phycoerythrin from Polysiphonia urceolata.

In SDS-PAGE gels of three purified R-phycoerythrins (R-PEs) isolated from three species of red algae, two bands whose molecular weights were about 40 kDa and 50 kDa can stably be found when the sample loading amount was enough. It is important for structure study of R-PE to clarify what these bands represent and how they are formed. According to results of the second SDS-PAGE, as well as molecular weights, fluorescences under UV and abundance, the 42.1 kDa and 53.7 kDa bands in SDS-PAGE gels of R-PE from Polysiphonia urceolata were believed to be complexes of αß and ßγ1, respectively. Formation of these bands may be related to light and phycourobilins (PUB) in subunits; and appearance of these two bands provided some proofs on position of chromophores and directions of energy transfer in R-PE. R-PE containing γ1 subunit was obviously more stable than R-PE containing γ2 subunit when they were exposed to protein denaturants, so γ subunits of R-PE may play important roles in structural stability of R-PE aggregates and the main forces that maintain the stability of R-PE may be interactions between γ subunit and ß subunits.

Renal transplantations from parents to siblings with autosomal recessive Alport syndrome caused by a rearrangement in an intronic antisense Alu element in the COL4A3 gene led to different outcomes.

Two siblings with autosomal recessive Alport syndrome (ARAS) obtained renal transplants from their consanguineous parents. Their COL4A3 mRNA transcripts were disrupted by a 139 bp intronic sequence between exon 48 and 49, which was derived from an antisense Alu element in this intron. The new amino acid sequence from the cryptic exon was terminated by a stop codon at the 1511th codon, resulting in the loss of 76 % α3(IV)NC1. This is the first case report of kidney transplantations between ARAS-homozygous siblings and their heterozygous parents. The brother experienced acute rejection just after transplantation and post-transplantation anti-glomerular basement membrane (GBM) nephritis, whereas the sister has experienced no problems to date. The anti-GBM nephritis could have resulted from the acute rejection. The COL4A3 gene heterozygous mutated parents, who are possibly at risk for thin basement membrane disease, have maintained their renal functions without urinary abnormalities after renal transplantation to date.