1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
use crate::cli::CenterOptions;

use super::fiber::FiberseqData;
use super::*;
use bamlift::*;
use bio::alphabets::dna::revcomp;
use bio_io;
use indicatif::{style, ProgressBar};
use rayon::prelude::*;
use rust_htslib::bam::Read;
use rust_htslib::{bam, bam::ext::BamRecordExtensions};
use std::fmt::Write;
use std::io::{self, prelude::*};

#[derive(Clone, Debug, PartialEq, Eq)]
pub struct CenterPosition {
    pub chrom: String,
    pub position: i64,
    pub strand: char,
}
pub struct CenteredFiberData {
    fiber: FiberseqData,
    pub dist: Option<i64>,
    center_position: CenterPosition,
    pub offset: i64,
    pub reference: bool,
    pub simplify: bool,
}

impl CenteredFiberData {
    pub fn new(
        fiber: FiberseqData,
        center_position: CenterPosition,
        dist: Option<i64>,
        reference: bool,
        simplify: bool,
    ) -> Option<Self> {
        let (ref_offset, mol_offset) =
            CenteredFiberData::find_offsets(&fiber.record, &center_position);
        let offset = if reference { ref_offset } else { mol_offset };

        let fiber = fiber.center(&center_position)?;

        Some(CenteredFiberData {
            fiber,
            dist,
            center_position,
            offset,
            reference,
            simplify,
        })
    }
    /// find both the ref and mol offsets
    pub fn find_offsets(record: &bam::Record, center_position: &CenterPosition) -> (i64, i64) {
        let ref_offset = center_position.position;
        let mol_offset =
            CenteredFiberData::find_offset(record, center_position.position).unwrap_or(0);
        (ref_offset, mol_offset)
    }

    /// find the query position that corresponds to the central reference position
    pub fn find_offset(record: &bam::Record, reference_position: i64) -> Option<i64> {
        let read_center: Vec<i64> = lift_query_positions_exact(record, &[reference_position])
            .into_iter()
            .flatten()
            .collect();
        log::debug!(
            "{}, {}, {}, {:?}",
            reference_position,
            record.reference_start(),
            record.reference_end(),
            read_center
        );
        if read_center.is_empty() {
            None
        } else {
            Some(read_center[0])
        }
    }

    /// Get the sequence
    pub fn subset_sequence(&self) -> String {
        if self.simplify {
            return "N".to_string();
        }
        let dist = if let Some(dist) = self.dist { dist } else { 0 };
        let seq = self.fiber.record.seq().as_bytes();

        let mut out_seq: Vec<u8> = vec![];
        let st = self.offset - dist; //(self.offset - dist)
        for pos in st..(self.offset + dist + 1) {
            if pos < 0 || pos as usize >= seq.len() {
                out_seq.push(b'N');
            } else {
                out_seq.push(seq[pos as usize]);
            }
        }
        if self.center_position.strand == '-' {
            out_seq = revcomp(out_seq);
        }
        //assert_eq!(out_seq.len() as i64, dist * 2 + 1);
        String::from_utf8_lossy(&out_seq).to_string()
    }

    pub fn get_sequence(&self) -> String {
        let forward_bases = if self.center_position.strand == '+' {
            self.fiber.record.seq().as_bytes()
        } else {
            revcomp(self.fiber.record.seq().as_bytes())
        };
        String::from_utf8_lossy(&forward_bases).to_string()
    }

    pub fn leading_columns(&self) -> String {
        let (mut c_query_start, mut c_query_end) = if self.reference {
            (
                self.fiber.record.reference_start() - self.center_position.position,
                self.fiber.record.reference_end() - self.center_position.position,
            )
        } else {
            let query_length = self.fiber.record.seq_len() as i64;
            (-self.offset, query_length - self.offset)
        };

        if self.center_position.strand == '-' {
            c_query_start = -c_query_start;
            c_query_end = -c_query_end;
            if c_query_start > c_query_end {
                std::mem::swap(&mut c_query_start, &mut c_query_end);
            }
        }
        format!(
            "{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t",
            self.center_position.chrom,
            self.center_position.position,
            self.center_position.strand,
            self.subset_sequence(),
            self.fiber.record.reference_start(),
            self.fiber.record.reference_end(),
            std::str::from_utf8(self.fiber.record.qname()).unwrap(),
            self.fiber.rg,
            self.fiber.get_hp(),
            c_query_start,
            c_query_end,
            self.fiber.record.seq_len()
        )
    }

    #[allow(clippy::type_complexity)]
    fn grab_data(
        &self,
    ) -> (
        &[Option<i64>],
        &[u8],
        &[Option<i64>],
        &[u8],
        &[Option<i64>],
        &[Option<i64>],
        &[Option<i64>],
        &[Option<i64>],
        &[u8],
    ) {
        if self.reference {
            (
                &self.fiber.m6a.reference_starts,
                &self.fiber.m6a.qual,
                &self.fiber.cpg.reference_starts,
                &self.fiber.cpg.qual,
                &self.fiber.nuc.reference_starts,
                &self.fiber.nuc.reference_ends,
                &self.fiber.msp.reference_starts,
                &self.fiber.msp.reference_ends,
                &self.fiber.msp.qual,
            )
        } else {
            (
                &self.fiber.m6a.starts,
                &self.fiber.m6a.qual,
                &self.fiber.cpg.starts,
                &self.fiber.cpg.qual,
                &self.fiber.nuc.starts,
                &self.fiber.nuc.ends,
                &self.fiber.msp.starts,
                &self.fiber.msp.ends,
                &self.fiber.msp.qual,
            )
        }
    }

    pub fn write(&self) -> String {
        let (m6a, m6a_qual, cpg, cpg_qual, nuc_st, nuc_en, msp_st, msp_en, fire) = self.grab_data();
        format!(
            "{}{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\n",
            self.leading_columns(),
            join_by_str_option(m6a, ","),
            join_by_str(m6a_qual, ","),
            join_by_str_option(cpg, ","),
            join_by_str(cpg_qual, ","),
            join_by_str_option(nuc_st, ","),
            join_by_str_option(nuc_en, ","),
            join_by_str_option(msp_st, ","),
            join_by_str_option(msp_en, ","),
            join_by_str(fire, ","),
            self.get_sequence(),
        )
    }

    pub fn leading_header() -> String {
        format!(
            "{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t",
            "chrom",
            "centering_position",
            "strand",
            "subset_sequence",
            "reference_start",
            "reference_end",
            "query_name",
            "RG",
            "HP",
            "centered_query_start",
            "centered_query_end",
            "query_length",
        )
    }
    pub fn header() -> String {
        format!(
            "{}{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\t{}\n",
            CenteredFiberData::leading_header(),
            "centered_m6a_positions",
            "m6a_qual",
            "centered_5mC_positions",
            "5mC_qual",
            "centered_nuc_starts",
            "centered_nuc_ends",
            "centered_msp_starts",
            "centered_msp_ends",
            "query_sequence"
        )
    }

    pub fn long_header() -> String {
        format!(
            "{}{}\t{}\t{}\t{}\n",
            CenteredFiberData::leading_header(),
            "centered_position_type",
            "centered_start",
            "centered_end",
            "centered_qual",
        )
    }

    pub fn write_long(&self) -> String {
        let mut rtn = String::new();
        let (m6a, m6a_qual, cpg, cpg_qual, nuc_st, nuc_en, msp_st, msp_en, fire) = self.grab_data();
        for (t, vals) in [
            ("m6a", (m6a, None, Some(m6a_qual))),
            ("5mC", (cpg, None, Some(cpg_qual))),
            ("nuc", (nuc_st, Some(nuc_en), None)),
            ("msp", (msp_st, Some(msp_en), Some(fire))),
        ] {
            let starts = vals.0.iter().collect::<Vec<_>>();
            let ends: Vec<Option<i64>> = match vals.1 {
                Some(ends) => ends.to_vec(),
                None => vec![None; starts.len()],
            };
            let quals = match vals.2 {
                Some(quals) => quals.to_vec(),
                None => vec![0; starts.len()],
            };

            for ((&st, &en), &qual) in starts.iter().zip(ends.iter()).zip(quals.iter()) {
                let Some(st) = st else {
                    continue;
                };
                let st = *st;
                let en = en.unwrap_or(st + 1);

                let mut write = true;
                if let Some(dist) = self.dist {
                    // skip writing if we are outside the motif range
                    if en <= -dist || st > dist {
                        write = false;
                    }
                };
                if write {
                    // add the leading data
                    rtn.push_str(&self.leading_columns());
                    // add the long form data
                    write!(
                        &mut rtn,
                        "{}",
                        format_args!("{}\t{}\t{}\t{}\n", t, st, en, qual)
                    )
                    .unwrap();
                }
            }
        }

        rtn
    }
}

#[allow(clippy::too_many_arguments)]
pub fn center(
    records: Vec<bam::Record>,
    header_view: &rust_htslib::bam::HeaderView,
    center_position: CenterPosition,
    min_ml_score: u8,
    wide: bool,
    dist: Option<i64>,
    reference: bool,
    simplify: bool,
    buffer: &mut Box<dyn std::io::Write>,
) {
    let fiber_data = FiberseqData::from_records(records, header_view, min_ml_score);
    let total = fiber_data.len();
    let mut seen = 0;

    fiber_data
        .into_par_iter()
        .map(|fiber| {
            match CenteredFiberData::new(fiber, center_position.clone(), dist, reference, simplify)
            {
                Some(centered_fiber) => {
                    if wide {
                        centered_fiber.write()
                    } else {
                        centered_fiber.write_long()
                    }
                }
                None => "".to_string(),
            }
        })
        .collect::<Vec<_>>()
        .iter()
        .filter(|x| !x.is_empty())
        .for_each(|x| {
            seen += 1;
            write_to_file(x, buffer);
        });

    if total - seen > 1 {
        log::warn!(
            "Unable to exactly map {}/{} reads at position {}:{}",
            total - seen,
            total,
            center_position.chrom.clone(),
            center_position.position
        );
    }
}

pub fn center_fiberdata(
    center_opts: &CenterOptions,
    bam: &mut bam::IndexedReader,
) -> anyhow::Result<()> {
    let center_positions = center::read_center_positions(&center_opts.bed)?;

    // header needed for the contig name...
    let header = bam::Header::from_template(bam.header());
    let header_view = bam::HeaderView::from_header(&header);
    //
    let mut buffer = bio_io::writer("-").unwrap();

    if center_opts.wide {
        bio_io::write_to_file(&CenteredFiberData::header(), &mut buffer);
    } else {
        bio_io::write_to_file(&CenteredFiberData::long_header(), &mut buffer);
    }

    let pb = ProgressBar::new(center_positions.len() as u64);
    pb.set_style(
        style::ProgressStyle::with_template(PROGRESS_STYLE)
            .unwrap()
            .progress_chars("##-"),
    );

    for center_position in center_positions {
        bam.fetch((
            &center_position.chrom,
            center_position.position,
            center_position.position + 1,
        ))
        .unwrap_or_else(|_| {
            panic!(
                "Failed to fetch region: {}:{}-{}",
                &center_position.chrom,
                center_position.position,
                center_position.position + 1
            )
        });
        let records: Vec<bam::Record> = bam.records().map(|r| r.unwrap()).collect();
        center(
            records,
            &header_view,
            center_position,
            center_opts.min_ml_score,
            center_opts.wide,
            center_opts.dist,
            center_opts.reference,
            center_opts.simplify,
            &mut buffer,
        );
        pb.inc(1);
    }
    buffer.flush().unwrap();
    pb.finish_with_message("\ndone");
    Ok(())
}

pub fn read_center_positions(infile: &str) -> io::Result<Vec<CenterPosition>> {
    let reader = bio_io::buffer_from(infile).expect("Failed to open bed file");
    let mut rtn = vec![];
    for line in reader.lines() {
        let line = line?;
        if line.starts_with('#') {
            continue;
        }
        let tokens = line.split('\t').collect::<Vec<_>>();
        assert!(tokens.len() >= 3);
        let st = tokens[1].parse::<i64>().unwrap();
        let en = tokens[2].parse::<i64>().unwrap();
        // get the strand for the 6 or 4th column
        let strand =
            if (tokens.len() >= 6 && tokens[5] == "-") || (tokens.len() >= 4 && tokens[3] == "-") {
                '-'
            } else {
                '+'
            };

        let (strand, position) = if strand == '-' {
            ('-', en - 1)
        } else {
            ('+', st)
        };
        rtn.push(CenterPosition {
            chrom: tokens[0].to_string(),
            position,
            strand,
        });
    }
    Ok(rtn)
}